Digital Fluid Teleportation, Advanced Biological Virtualization, And Large Scale Integration Of Organ-On-Chips And Microphysiological Models

ABSTRACT

A microphysiological platform described herein includes a fluidic synthesizer with a first fluid input selectively coupleable to a source of a first input fluid solution and a second fluid input selectively coupleable to a source of a second input fluid solution. The fluidic synthesizer further includes a fluid output. The microphysiological platform further includes a fluid addressing system with a fluid input fluidically coupled to the fluidic synthesizer fluid output. The fluid addressing system further includes a first fluid output and a second fluid output. The microphysiological platform further includes a first microphysiological device with a fluid input fluidically coupled to the first fluid output of the fluid addressing system and a second microphysiological device with a fluid input fluidically coupled to the second fluid output of the fluid addressing system.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the priority of U.S. Provisional PatentApplication Ser. No. 63/015,242, filed on Apr. 24, 2020, the disclosureof which is incorporated herein by reference.

BACKGROUND

Certain organ-on-chip devices can include microengineered biologicalcell-culture compartment in which tissue- and organ-level elements ofhuman physiology can be recapitulated. Cells implanted therein can beexpected to behave and organize in a physiologically realistic andrelevant manner, allowing accurate in vitro modeling of functionalbiological units of the organ system.

However, certain organ-chip systems fail to create functional and/orrealistic multi-organ networks and thus are unable to recapitulatecomplex, physiological responses and multi-organ interactions at thesystemic level. Each organ-chip can require a different composition ofgrowth media that needs to be formulated with hormones, nutrients, andother molecules to mimic the native microenvironment of the tissues inthat specific organ. Due to the different biochemical cues required bydifferent cell types to maintain their differentiated functions indifferent organs, it can be difficult to produce one single universalmedia composition that can be carried between organ units and stillsupport the appropriate differentiation and long-term maintenance of allthese multiple organ units simultaneously.

Furthermore the translation of organ-on-chip devices into practicalscreening platforms can face challenges in terms of the complexity ofoperating these devices. For example, fluids in certain organ-on-chipdevices need to be driven either by bulky syringe pumps or manually byscientists with pipettes. This reduced throughput can limit the adoptionof organ-on-chip models in the pharmaceutical industry because processeslike drug-candidate screening rely on automated, large-quantity, andhigh-throughput testing.

Accordingly, there remains a need to improve the throughput and theefficiency of the organ-on-chip devices. There is also a need forimproved organ-on-chip devices, which can permit certain transmissionsof cell-released signaling molecules, including hormones, between organsor tissues while relegating certain growth media formulations that candefine a certain tissue microenvironment solely to the organs for whichthey are intended.

SUMMARY

Embodiments described herein relate to microphysiological platforms andmethods of producing the same. In some embodiments, a microphysiologicalplatform can include fluidic synthesizer with a first fluid inputselectively coupleable to a source of a first input fluid solution and asecond fluid input selectively coupleable to a source of a second inputfluid solution. The fluidic synthesizer further includes a fluid output.The fluidic synthesizer creates an output solution by mixing first inputfluid solution received from the first fluid input and second inputfluid solution received from the second fluid input and discharges theoutput solution from the fluid output. The microphysiological platformfurther includes a fluid addressing system with a fluid inputfluidically coupled to the fluidic synthesizer fluid output. The fluidaddressing system further includes a first fluid output and a secondfluid output. The fluid addressing system conveys output solution fromthe fluid addressing system fluid input to a selected one, or both, ofthe first fluid output and the second fluid output Themicrophysiological platform further includes a first microphysiologicaldevice with a fluid input fluidically coupled to the first fluid outputof the fluid addressing system and a second microphysiological devicewith a fluid input fluidically coupled to the second fluid output of thefluid addressing system. Each of the first microphysiological device andthe second microphysiological device culture biological tissue andperfuse the biological tissue with the output solution from the fluidicsynthesizer received at the fluid input of the respectivemicrophysiological device via the fluid addressing system. In someembodiments, the fluidic synthesizer can have at least a third fluidinput selectively coupleable to at least a source of at least a thirdinput fluid solution and is further operable to create the inputsolution by mixing one or both a the first input fluid solution receivedfrom the first fluid input and the second input fluid solution receivedfrom the second fluid input the at least a third input fluid solutionreceived at the at least a third fluid input. In some embodiments, thefluidic synthesizer can include a mixing chamber fluidically coupled tothe fluid inputs and to the fluid output of the fluidic synthesizer andmix the input fluid solutions to create the output solution.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a schematic illustration of a microphysiological platform,according to an embodiment.

FIG. 2A is a schematic illustration of a control system, and FIG. 2B isa schematic illustration of the microphysiological platform of FIG. 1engaged with a control system, according to an embodiment.

FIG. 3 is schematic illustration of the fluidic synthesizer of themicrophysiological platform of FIG. 1 , according to an embodiment.

FIG. 4 is a schematic illustration of the fluid addressing system of themicrophysiological platform of FIG. 1 , according to an embodiment.

FIGS. 5A to 5C are each a schematic illustration of an embodiment of amicrophysiological device that may be included in the microphysiologicalplatform of FIG. 1 . FIG. 5B is a schematic illustration of amicrophysiological device in a horizontal configuration, and FIG. 5C isa schematic illustration of a microphysiological device in a verticalconfiguration.

FIG. 6A is a schematic top view, and FIG. 6B is a schematic sidecross-sectional view of a microphysiological device and biosensorarrangement, according to an embodiment, FIG. 6C is an exemplaryillustration of an organ tissue culture contained in themicrophysiological device of FIGS. 6A and 6B, and FIG. 6D is a detailedschematic top view of the biosensor of FIGS. 6A and 6B.

FIG. 7A is a schematic illustration of the augmentation of discretemicrophysiological devices with on-chip synthesizers and FIG. 7B is aschematic illustration of discrete microphysiological devices withon-chip fluidic synthesizers integrated into a single monolithicmicrophysiological platform.

FIG. 8A shows an illustration of a microphysiological platform thatintegrates numerous individual microphysiological devices withassociated sensors in a single multilayer microfluidic device, accordingto an embodiment. FIGS. 8B, 8C, and 8D show portions of themicrophysiological platform.

FIG. 9 is a schematic illustration of a fluidic teleportation systemaccording to an embodiment.

FIG. 10 is a schematic illustration of a fluidic teleportation system,according to an embodiment.

FIGS. 11A to 11D are schematic illustrations of different topologies offluidic teleportation systems, according to various embodiments.

FIG. 12 is a schematic illustration of a human-body-on-a-chip modelincluding multiple microphysiological devices that each recapitulate thephysiology or function of a particular human tissue, organ, or system,each connected to one or more other devices by digital fluidicteleportation.

FIG. 13 is a flow diagram illustrating a method of fluidicteleportation, according to an embodiment.

FIG. 14 is an illustration of an exemplary system hardware architectureof a microphysiological system including multiple microphysiologicalplatforms, according to an embodiment.

FIGS. 15A and 15B are flow diagrams illustrating a method of ascreen-forward mode of fluid teleportation, according to an embodiment.

FIGS. 16A-16D are flow diagrams illustrating a method of ascreen-backward mode of fluid transportation, according to anembodiment.

FIG. 17A is an illustration of an exemplary user interface view ofsystem software, and FIG. 17B is an illustration of exemplarymicrophysiological device interconnection visualization, used in thisexemplary embodiment to illustrate active fluid teleportationconnections, according to an embodiment.

FIG. 18 is an illustration of exemplary data collection using a virtualtissue in accordance with the present disclosure.

FIG. 19A is an illustration of an exemplary coupling of amicrophysiological device—containing a fluidic synthesizer, a biologicalculture chamber, and a biosensor—with another microphysiological device.FIG. 19B s an illustration of an exemplary coupling of a singlemicrophysiological device to a dynamic system of multiple virtualtissues, using an exemplary embodiments of digital fluid teleportationas an interconnect in accordance with the present disclosure, FIG. 19Cis an illustration of an exemplary interface of multiple virtual tissuestogether by an exemplary usage of digital fluid teleportation to form afully virtual, according to various embodiments.

FIG. 20 is a schematic illustration of a controller such as a maincontroller of a control system, according to an embodiment.

Throughout the figures, the same reference numerals and characters,unless otherwise stated, are used to denote like features, elements,components or portions of the illustrated embodiments. Moreover, whilethe present disclosure will now be described in detail reference to thefigures, it is done so in connection with the illustrative embodiments.

DETAILED DESCRIPTION

The disclosed subject matter provides systems and methods for one or acombination a integration, management, automation, analoginterconnection, digital interconnection, maintenance, observation,analysis, or electronic control, of one or a plurality ofmicrophysiological devices.

Techniques for producing a microphysiological platform including one ora plurality of microphysiological devices are disclosed herein. Thedisclosed subject matter can perform a fully or partially automatedbiological culture using the microphysiological devices without the needfor specialized personnel. In certain embodiments, it can model thefeed-forward and feed-back effects between biological systems orbiological tissues modeled in a plurality of microphysiological devices,among which one or multiple subsets can be interconnected.

Microphysiological Platform and Control System Architecture

FIG. 1 is a schematic illustration of a microphysiological platform 100,according to an embodiment. As shown in FIG. 1 microphysiologicalplatform 100 can include a synthesizer 110, a fluid addressing system120, an array of microphysiological devices 130, an army o biosensors140, a sensor data transmitter 150, and an effluent channel 160. Fluidscan flow into platform 100, such as into fluid inputs 112 of fluidicsynthesizer 110, through and between the components, and out of platform100, such as out of fluid output 164 of effluent channel 160. Suchfluids can include a wash fluid and one or more input solutions. Asdescribed in more detail below with reference to FIG. 3 , multiple inputsolutions received at fluidic inputs 112 of fluidic synthesizer 110 canbe mixed within fluidic synthesizer 110, in some embodiments under thecontrol of control inputs received by fluidic synthesizer 110, and theresultant customized solution can be output from fluid output 114. Fluidoutput 114 is fluidically coupled to fluid input 122 of fluid addressingsystem 120. As described below with reference to FIG. 4 , fluidaddressing system 120 can route a discrete bolus, or continuous flow, ofthe output solution from fluid input 122 to a selected one of fluidoutputs 124. The routing of the fluid can be controlled by controlinputs received by fluid addressing system 120. Each fluid output 124 isfluidically connected to an input of a specific one ofmicrophysiological devices 130 in the array microphysiological devices130. As described in more detail below with reference to FIGS. 5A to 5C,each microphysiological device 130 may be implemented as an organ on achip, and the solution delivered to the input of microphysiologicaldevice 130 can interact with the content of the device 130, and itscomposition changed before reaching the output of device 130. The outputof device 130 is fluidically coupled to the input of one or morebiosensors 140 in the array of biosensors 140, such that the bolus ofmodified fluid can be received by one or more of the biosensors 140. Asdescribed in more detail below in part with reference to FIGS. 6A to 6D,each biosensor 140 can detect of measure one or more properties of themodified solution, and can generate a data signal indicative of the oneor more properties. In some embodiments the generated sensor data can becommunicated to a sensor data transmitter 150, which can transmit thesensor data through a data output 154 to be received by a data receiverseparate from the microphysiological platform 100. In some embodiments,the generated sensor data can be read directly from the biosensors 140(i.e., with the use of the sensor data transmitter 150). In other words,the generated sensor data can be read without the use of a datatransmission system. Each biosensor 140 has a fluid output that isfluidically connected to an input of a effluent channel 160 such thatthe modified fluid can he conveyed from biosensor 140 to effluentchannel 160, from which it can be discharged through fluid output 164 ofeffluent channel 160 to a effluent separate from the microphysiologicalplatform 100. In some embodiments, the fluid output 164 can be routed torecycle fluid. In some embodiments, the fluid output 164 can route fluidback to the fluid addressing system 120. In some embodiment, the fluidoutput 164 can route fluid back to fluidic synthesizer 110. In someembodiments, the fluid output 164 can route fluid back to the array ofmicrophysiological devices 130. In some embodiments, the fluid output164 can undergo further processing prior to being re-integrated into themicrophysiological platform 100.

In some embodiments, the fluidic synthesizer 110 can generate microliterquantities of a specified fluidic mixture to perform conditional orcombinatorial screening on all or some of the microphysiological devices130 m the microphysiological platform 100. In some embodiments, thefluidic synthesizer 110 can generate quantities ranging from less thannanoliter to more than one milliliter of a specified fluidic mixture. Insome embodiments, the quantities generated of one or a plurality offluid mixtures can depend on the requirements of a specific platform, anintended experiment, a biological system, or a combination thereof.

In some embodiments, the fluid addressing system in can be controlledmanually, semi-automatically or automatically to dynamically changewhich individual microphysiological devices or which subset ofmicrophysiological devices is selected as outputs for delivery of aninput fluid or fluids. In certain embodiments, the fluid addressingsystem 120 can include at least one value that is configured to divertthe flow of a fluidic mixture to a plurality of selected outputs ormicrophysiological devices 130.

In some embodiments the fluidic synthesizer 110 can repeatedly producethe at least one target fluid mixture which can be delivered by thefluidic addressing system 120 to a plurality of microphysiologicaldevices 130. In non-limiting embodiments, such operation can be cycledautonomously to create an automated microphysiological platform.

In some embodiments, the microphysiological platform 100 can include afirst fluidic synthesizer 110 for creating a first fluidic mixture, afirst fluid addressing system 120 for routing first fluid mixture to allor a subset of at least one microphysiological device 130, and at leastone microphysiological device 130 for incubating a biological tissue ormaterial with the first fluidic mixture to generate a second fluidicmixture. For example, the second fluidic mixture can be generated bybiochemical interaction between the first fluidic mixture and the firstmicrophysiological device.

In some embodiments, the biologic tissue can include a lung tissue, abone marrow tissue, a bone tissue, a pancreatic tissue, an endocrineislets tissue, a liver tissue, a kidney tissue a placenta tissue an eyetissue, an intestinal tissue, a bladder tissue, a brain tissue, a mouthtissue, a tongue tissue, a tooth tissue, a nose tissue, a thymus tissue,a lymph node tissue, a lymphatic system tissue, a throat tissue, or amcombination thereof. In some embodiments, the biologic tissue caninclude a specific human tissue. In some embodiments, a specific humantissue can include one or more of a human organ, an organ subcomponent,a system of two or more organs, a system of a generic tissue element(e.g., blood vessel or a ligament), or a specific cell type (e.g., afibroblast cell that might be engaging in behavior of interest such ascreating fibrosis). In some embodiments, the tissue can include aspecific human tissue undergoing a specific routine behavior, a lung,tissue that is cyclically breathing, a specific human tissue undergoing,an atypical condition, a lung tissue undergoing an asthma attack, aspecific human tissue undergoing a specific interaction with an outsideagent a lung tissue being infected with bacteria, a lung tissue exposedto environmental factors, a lung tissue exposed to pollution, a longtissue exposed to corrosive gas specific human tissue undergoing aspecific interaction with an outside agent that is intended for use as atherapeutic, a specific human tissue undergoing a specific interactionwith a drug, a specific human tissue undergoing a specific interactionwith a biological antibody, a specific human tissue undergoing aspecific interaction with a cellular therapy, a lung tissue undergoingan asthma attack while being monitored for its interaction with abronchodilator as therapy for asthma, or any combination thereof.

In some embodiments, the biosensors 140 can include at least onechemical sensor. In some embodiments, the at least one chemical sensorcan be coupled to the at least one microphysiological device 130 fordetecting target analytes in the second fluidic mixture. In someembodiments, the at least one chemical sensor can include a thinconductive film for generating a resonant plasmonic coupling withincident light, and at least one biorecognition molecule for binding tothe at least one target analyte to create an SPR sensor. The thin filmcan be configured to induce a transducable disturbance or shift in saidresonant coupling when the at least one biorecognition molecule binds tothe target can include a transducer for transmitting a measured targetanalyte data (e.g., name and concentration of target analyte) to anexternal receiver. In some embodiments, the microphysiological platform100 can include at least one external receiver, which can be configuredto monitor and/or process the measured target analyte data. In someembodiments, the at least one sensor can utilize at least one sensingmodality in addition to the SPR chemical sensing.

Microphysiological platform 100 can be supported by, can receive fluidand control inputs from, and can provide data and fluid outputs to, acontrol system 170, shown schematically in FIG. 2A. As shown in FIG. 2A,control system 170 can include controllers, a data system 180, andengagement system 185, and a fluid system 190. Controllers can include amain controller 172, one Or more fluid synthesizer controllers 174, oneor more addressing system controllers 176, and an external communicator178. Data system 180 can include an imager 182 and one or more sensordata receivers 181. Engagement system 185 can include a mechanicalsupport 186, one or more fluid couplings 187, and one or more controlline couplings 188. Fluid system 190 can include one or more inputsolution reservoirs 192, one or more wash solution reservoirs 194, oneor more fluid handlers 195, and one or more effluent reservoirs 196.

For example, the control system 170 can include fluid reservoirs, suchas a wash fluid reservoir 194 and reservoirs for input solutions for thefluidic synthesizer 110, and/or can provide fluid conduits tofluidically couple reservoirs external to the control system 170, toprovide fluids in the respective inputs on the microphysiologicalplatform 100. It can also provide a reservoir for fluid output 164 fromthe effluent channel 160 of the microphysiological platform 100, and/orcan provide fluid conduit(s) to fluidically couple effluent reservoir(s)external to the control system 170. The control system 170 can alsoinclude one or more controllers, such a fluidic synthesizer controller174 to control the operation of the fluidic synthesizer 110 (e.g.control application of pneumatic pressure to valves in the fluidicsynthesizer 110), and an addressing system controller 176 to control theoperation of the fluid addressing system 120 (e.g. via communicationwith valves in the fluid addressing system 120 to guide fluid flow) Thecontrol system 170 can include a data receiver 181 to receive sensordata provided by the sensor data transmitter(s) 150 associated with thebiosensors. It can also include an imager 182 (such as a charge-coupleddevice, an active-pixel sensor, an optical microscope, a magneticresonance imaging machine, a computed tomography machine or a MOSfield-effect transistor sensor) that can acquire image data from themicrophysiological devices 130 and/or biosensors 140. The control system170 can also include a main controller to receive data from the othercontrollers and components, and/or provide instructions thereto, and cancommunicate with other devices or systems external to the control system170.

In some embodiments, the fluid handler 195 can selectively deliver fluiddirectly to desired locations on the microphysiological platformincluding any one of the microphysiological devices 130 in the array ofmicrophysiological devices 130. In other words, each of themicrophysiological devices 130 can receive fluid from the fluidaddressing system 120 as well as from the fluid handler 195. In someembodiments, the fluid handler 195 can include one or moremicroinjectors and/or one or more pipettes. In some embodiments, thefluid handler 195 can be movable, such that the fluid handler 195 candeliver fluid to a specified microphysiological device 130 among thearray of microphysiological devices 130. This direct delivery can limitfluidic loss. For example, if fluid A is being delivered to a specifiedmicrophysiological device 130 and the user needs to deliver fluid B tothe microphysiological device 130, the user can do so without flushingfluid A from the path in the fluid addressing system 120 and the fluidicsynthesizer 110. This direct delivery capability can also betime-efficient, as the user does net need to take the time to flush thelines of fluid A before delivering fluid B to a desired site. In someembodiments, the fluid handler 195 can couple to the same ports on themicrophysiological devices 130 as the fluid addressing system 120. Insome embodiments, the fluid addressing system 120 can couple to a firstport on a microphysiological device 130 and the fluid handler 195 cancouple to a second port on the microphysiological device 130.

In some embodiments, the fluid handler 195 can deliver fluid to alocation upstream of the microphysiological devices 130 (e.g., upstreamof an interface between the fluid addressing system 120 and themicrophysiological devices 130). In some embodiments, the fluid handler195 can deliver fluid in-line to the microphysiological devices 130. Inother words, a port can be integrated into a section of one or moremicrophysiological devices 130 for access by the fluid handler 195. Insome embodiments, fluid handler 136 can deliver fluid to a locationdownstream of the microphysiological devices, 130. In some embodiments,each of the microphysiological devices 130 can include a valve forinjection from the fluid hander 195.

In some embodiments, the fluid handler 195 can draw fluid from one ormore of the microphysiological devices 130. In some embodiments, thefluid handler 195 can withdraw fluid from a location upstream of themicrophysiological devices 130. In some embodiments, the fluid handler195 can draw fluid from a location in-line to the microphysiologicaldevices 130. In some embodiments, the fluid handler 195 can withdrawfluid from a location downstream of the microphysiological devices 130.

In some embodiments, the fluid handier 195 can deliver tissue, a tissueprogenitor, stem cells, or any combination thereof to themicrophysiological devices 130. In some embodiments, the fluid handler195 can be used to develop tissue on site in the microphysiologicaldevices 130. In some embodiments, tissue can be developed in the fluidhandler 195.

In some embodiments the fluid handler 195 can deliver or withdraw fluidfrom any portion of the microphysiological platform 100. In someembodiments, the fluid handler 195 can deliver fluid to the fluidicsynthesizer 110. In some embodiments, the fluid handler 195 can withdrawfluid from the fluidic synthesizer 110. In some embodiments, the fluidhandler 195 can deliver fluid to the fluid addressing system 120. Insome embodiments, the fluid handler 195 can withdraw fluid from thefluid addressing system 120. In some embodiments, the fluid handler 195can deliver fluid to the microphysiological devices 130. In someembodiments, the fluid handler 195 can withdraw fluid from themicrophysiological devices 130.

FIG. 2B is a schematic illustration of one microphysiological platform100 engaged with control system 170. As shown the mechanical support 186positions the substrate 101 to be in contact with one or more of thecomponents of the control system 170. In some embodiments, themechanical support 186 can include a clamp to engage the substrate 101and hold the substrate 101 in an operative relationship with one or morecomponents of the control system 170. For example, the mechanicalsupport 186 can support the substrate 101 in a read/write drive. In someembodiments, the mechanical support 186 can include structuralcomponents to hold the control line couplings 188 and the fluidcouplings 187 in place. For example, the mechanical support 186 caninclude one or more platforms or clamps to hold the control linecouplings 188 and the fluid couplings 187 in place. In some embodiments,the mechanical support 186 can act as, or include, a manifold forfluidic couplings (e.g., between the fluidic synthesizer 110 and thefluid addressing system 120). In some embodiments, the mechanicalsupport 186 can align the substrate 101 with a known position. In someembodiments, the position of the substrate 101 can be constrained suchthat one or more fiduciary pins (not shown) on the mechanical support186, whose position is known to the positioning systems in the fluidhandler 195 and the imager 182, can act as a datum against which toreference the positions of different items. In some embodiments, theseitems can include tissues, targets of interest on the substrate 101,and/or the substrate 101 itself.

In same embodiments, the fluid couplings 187 can fluidically couple thewash reservoir 194, the input solution reservoir 192, and the effluentreservoir 196 to the fluidic synthesizer 110, fluid addressing system120, and the microphysiological devices 130. In some embodiments, thefluid couplings 187 can be established with inputs on the components ofthe microphysiological platform 100 (e.g., inputs into the fluidicsynthesizer 110). In some embodiments the fluid couplings 187 can beestablished with outputs from the microphysiological platform 100 (e.g.,the outputs from the effluent channel 160). In some embodiments thecontrol line couplings 188 can be established with inputs on thecomponents of the microphysiological platform 100 (e.g., inputs into thefluidic synthesizer 110). In some embodiments, the control linecouplings 188 can be established with outputs from themicrophysiological platform 100 (e.g., the outputs from the effluentchannel 160).

In some embodiments, the data receiver 181 can establish communicationwith the sensor data transmitter 150. In some embodiments, the datareceived 181 can be operatively disposed to take passive readings fromthe biosensors 140.

In some embodiments, the imager 182 can be operatively disposed to imagethe components of the microphysiological platform . In some embodiments,the imager 182 can use light imaging techniques to detect fluids invarious portions of the fluidic synthesizer 110, the fluid addressingsystem 120, and/or the microphysiological devices 130. In someembodiments, the imager 182 can be used to detect a type of cell or cellculture in the microphysiological devices 130 based on how light isabsorbed, reflected, or scattered from the microphysiological devices130.

In some embodiments, the main controller 172 can control the fluidicsynthesizer controller 174, the addressing system controller 176, thedata receiver 181, and the imager 182. In some embodiments, the fluidicsynthesizer controller 174 can control the valve openings and the fluidflow rates into and/or out of the fluidic synthesizer 110 (i.e., thecontrol line couplings 188). For example, the user can specify aspecific mixture ratio and flow rate that the user wants to deliver tothe fluid addressing system 120 via the fluidic synthesizer controller174, and the appropriate valves can be opened and closed in the fluidicsynthesizer 110 via the fluidic synthesizer controller 174.

In some embodiments, the addressing system controller 176 can controlthe valve openings and the fluid flow rates into and/or out of the fluidaddressing system 120. In some embodiments, the addressing systemcontroller 176 can control the opening and closing of valves between thefluid addressing system 120 and the microphysiological devices 130. Insome embodiments, the addressing system controller 176 can controlpressures in the fluid addressing system 120 and the microphysiologicaldevices 130 via controlling the valves in the fluid addressing system120.

In some embodiments, the fluidic synthesizer controller 174, theaddressing system controller 176, the imager 182, and the data receiver181 can be monitored and/or controlled via a single user interface(i.e., via the main controller 172). In some embodiments, the fluidicsynthesizer controller 174, the addressing system controller 176, theimager 182, and the data receiver 181 can be monitored via separate userinterfaces. In some embodiments, the main controller 172 can communicatedata to the user via the external communicator 178.

In some embodiments, the main controller 172, the fluidic synthesizercontroller 174, and/or the addressing system controller 176 can includea proportional controller, an integral controller, a derivativecontroller, a proportional integral derivative (PID) controller, or anycombination thereof. Any of the controllers described above can beimplemented as general purpose compute device. For example, maincontroller 172 is shown schematically in FIG. 20 , and can include aprocessor 172A, and memory 172B, and one or more input/output devices172C. The processor 172A may be configured to receive, process, analyze,compile, store, and access data, e.g. through a network connection, asdiscussed in more detail herein, or through a physical connection withthe device or storage medium (e.g. through Universal Serial Bus (USB) orany other type of port). Processor 172A may be any suitable processingdevice configured to run and/or execute a set of instructions or codeand may include one or more data processors, image processors, graphicsprocessing units, physics processing units, digital signal processors,and/or central processing units. Each processor 172A may be for example,a general purpose processor, Field Programmable Gate Array (FPGA), anApplication Specific Integrated Circuit (ASIC), and/or the like. Eachprocessor 172A may be configured to run and/or execute applicationprocesses and/or other modules, processes and/or functions associatedwith the system and/or a network associated therewith. The underlyingdevice technologies may be provided in a variety of component types(e.g., metal-oxide semiconductor field-effect transistor (MOSFET)technologies like complementary metal-oxide semiconductor (CMOS),bipolar technologies like, emitter-coupled logic (ECL), polymertechnologies (e.g., silicon-conjugated polymer and metal-conjugatedpolymer-metal structures), mixed analog and digital, and/or the like.

Memory 172B may include a database (not shown) and may be, for example,a random access memory (RAM), a memory buffer, a hard drive, an erasableprogrammable read-only memory (EPROM), an electrically erasableread-only memory (EEPROM), a read-only memory (ROM). Flash memory, andthe like. Memory 172B may store instructions to cause the processor toexecute modules, processes, and/or functions associated with thecommunication device, such as patient data processing, sensormeasurement, viral infection probability estimation, user device orpatient monitoring control, authentication, encryption, and/orcommunication. Some embodiments described herein relate to a computerstorage product with a non-transitory computer-readable medium (also maybe referred to as a non-transitory processor-readable medium) havinginstructions or computer code thereon for performing variouscomputer-implemented operations. The computer-readable medium (orprocessor-readable medium) is non-transitory in the sense that it doesnot include transitory propagating signals per se (e.g., a propagatingelectromagnetic wave carrying information on a transmission medium suchas space or a cable). The media and computer code (also may be referredto as code or algorithm) may be those designed and constructed for thespecific purpose or purposes.

Examples of non-transitory computer-readable media include, but are notlimited to, magnetic storage media such as hard disks, floppy disks, andmagnetic tape; optical storage media such as Compact Disc/Digital VideoDiscs (CD/DVDs); Compact Disc-Read Only Memories (CD-ROMs), andholographic devices; magneto-optical storage media such as opticaldisks; solid state storage devices such as a solid state drive (SSD) anda solid state hybrid drive (SSHD); carrier wave signal processingmodules; and hardware devices that are specially configured to store andexecute program code, such as Application-Specific Integrated Circuits(ASICs), Programmable Logic Devices (PLDs), Read-Only Memory (ROM), andRandom-Access Memory (RAM) devices. Other embodiments described hereinrelate to a computer program product, which may include, for example,the instructions and/or computer code disclosed herein.

Each input/output device 172C may be coupled with processor 172A and172B and can be configured to communicate with other devices (such asother controllers and components of control system 170, other suchcontrol systems, and other devices, such as via external communicator178). Each input/output device 172C may include a network interfaceconfigured to connect the compute device to another system (e.g.,Internet, remote server, database) by wired or wireless connection. Insome embodiments, the network interface may include a radiofrequency(RF) receiver, transmitter, and/or optical (e.g., infrared) receiver andtransmitter configured to communicate with one or more devices and/ornetworks.

Microphysiological platform 100, and the components thereof, can beimplemented on a chip or substrate 101. The substrate 101 can bemanufactured through a cleanroom nanofabrication and/or a lithographicprocess similar to that used in the microprocessor fabricationindustries. For example, when embedded into a single microfluidicsubstrate, the microphysiological platform can be fabricated by castinga thermoset polymer onto molds defined by negative photoresists, by hotembossing or stamping processes that can imprint the intendedmicrofluidic patterns onto a thermoplastic material, and/or by injectionmolding. In certain embodiments, the platform can be fabricated frommultiple bonded layers, which may be optically aligned, such as byutilizing complementary fiduciary markings. In certain embodiments, aplatform fabricated from two or more layers can be bonded by theapplication of heat, by a dispensed adhesive, by the application ofuncured thermoset plastic, including uncured polydimethylsiloxane(PDMS), which can subsequently cure to provide bonding force, and/or bythe application of force in the direction of layer stratification forcontaining enclosed fluids or biological compounds with the resultingpressure-based seal. In some embodiments, biosensors 140 can beintegrated into the substrate 101. In some embodiments, the biosensors140 can be located separate from the substrate 101, and/or maybeexternal to the substrate 101. In some embodiments, the biosensors 140can be integrated into a structure separate from the substrate 101.

In certain embodiments, the microphysiological platform 100 can includevarious geometric features. The geometric features can be patterned intothe microphysiological platform to affect the flow path of liquid. Forexample, the geometric features can include one or a combination of thefollowings: selectivity permeable membranes to restrict passage of somefluid compounds or phases and not others (e.g., selective gas or liquidpermeability), porous membranes, notches, pillars, edges to manipulateliquid by capillary pinning or surface tension trapping, micropillar,nanopillar arrays, surface metal deposition, thin wall features toenable elastomeric actuation, molded or mechanically punched holes totransport fluid between bonded layers, matching geometric features toenable mechanical alignment of at least two layers of a multilayereddevice by optical overlay or mechanical conformal fit, arrows orindicators for the purpose of indicating to a user a location, port, orzone of interest, or fiduciary marks to assist autofocus, stitching,motion, or positional homing routines imaging or automated analysis.

In some embodiments, microphysiological platform 100 can include amicrofluidic system to which at least one inlet connection and at leastone outlet connection can be formed. For example, as described in moredetail below, an inlet connection can be a fluid input to fluidicsynthesizer 110, a control input to fluid addressing system 120, a fluidinput to a component on the microphysiological platform such as amicrophysiological device 130, or a combination thereof. An outletconnection can be a waste output, a sampling output, or a combinationthereof. The disclosed system can include an external compressor togenerate regulated gas pressures. The gas pressures can drive fluid flowand gate the fluid addressing system 120, as described in more detailbelow.

FIG. 3 is a schematic illustration of fluidic synthesizer 110. Fluidsynthesizer 110 can mix together two or more input solutions atindividually specified ratios or proportions to create a custom fluidmixture, or output solution. As shown in FIG. 3 , fluidic synthesizer110 can include two or more fluid inputs 112, identified in FIG. 3 asfluid input 1 and fluid input N (where N can be 2 or more). In someembodiments, the fluid entering the fluidic synthesizer 110 via fluid Ninput 1 can be the same as the fluid entering the fluidic synthesizer110 via fluid input N or any of the other fluid inputs 112. In someembodiments, a first fluid can enter the fluidic synthesizer 110 via afirst fluid input 112 and a second fluid can enter the fluidicsynthesizer 110 via a second fluid input 112, the second fluid differentfrom the first fluid. In some embodiments, many different fluids (e.g.,at least about 3, at least about 4, at least about 5, at least about 6,at least about 7, at least about 8, at least about 9, at least about atleast about 20, at least about 30, at least about 40, at least about 50,at least about 60, at least about 70, at least about 80, at least about90, or at least about 100) can enter the fluidic synthesizer 110 via thefluid inputs 112. Each fluid input 112 is fluidically connected (e.g. byany suitable fluid conduit or channel) to a mixing chamber 116. Eachfluidic input can be selectively fluidically coupled to a respectivereservoir of an input solution (not part of microphysiological platform100, but optionally part of control system 170) to receive the inputsolution and convey it to mixing chamber 116. For example, a reservoirof input solution 1 is coupleable to fluid input 1, so that inputsolution 1 can be delivered via fluid input 1 to mixing chamber 116, anda reservoir of input solution N is coupleable to fluid input N, so thatinput solution N can be delivered by fluid input N to mixture chamber116, A desired quantity (or flow rate) of input solution 1 and a desiredquantity (or flow rate) of input solution N can thus be received inmixing chamber 116, and mixed together (e.g. passively or via an activemechanism such as an agitator) to form a desired quantity or bolus (orflow rate), of an output solution. Mixing chamber 116 is fluidicallycoupled to fluid output 114 of synthesizer 110, from which the outputsolution (a discrete bolus, or a continuous flow at a desired flow rate)can be conveyed to downstream component(s), e.g. to one or more of themicrophysiological devices 130, such as through fluid addressing system120.

Wash input 113 is fluidically coupled to mixing chamber 116, and isselectively fluidically coupleable to a reservoir of wash solution (notpart of microphysiological platform 100). After a desired quantity ofoutput solution has been delivered from fluidic synthesizer 110 viafluid output 114, a quantity of wash solution can be conveyed to mixingchamber 116, via wash input 113, and can expel or clear the outputsolution from mixing chamber 116 through fluid output 114 and downstreamcomponents. In some embodiments, the mixing chamber 116 can includeadditional outlet streams (not shown) for expelling waste recirculatingfluid back into the fluidic synthesizer 110.

The quantity (or flow rate) of each input solution delivered via therespective fluid input 112 can be controlled by the input solutionreservoir, e.g. by use of a pump controlling the flow of fluid from thereservoir, and/or a valve associated with the reservoir. Alternatively,the quantity (or flow rate) of any one of more of the input solutionsdelivered to mixing chamber 116 can be controlled by a separatemechanism, such as active valve included in the fluidic synthesizer 110.As shown in FIG. 3 , the fluidic coupling between each fluid input 112and mixing chamber 116 can include a valve 115. Valve 115 can be open,permitting flow of input solution to mixing chamber 116, or closed,preventing the flow of input solution. Each valve 115 can be controlledby a respective control input (not part of microphysiological platform100), such as the synthesizer controller 174 included in the controlsystem 170 as described above with reference to FIG. 2 . In someembodiments, the valves 115 can include check valves, ball valves, globevalves, plug valves, needle valves, butterfly valves, pinch valves, gatevalves, relief valves, or any combination thereof. In some embodiments,fluidic synthesizer 110 can be controlled by closed-loop controlsystems, including without limitation closed-loop fluidic control offlow rates by utilizing a feedback control loop (including, withoutlimitation, a proportional-integral-derivative controller or “PID”controller) between one or a combination of a flow sensor, a pressureregulator, and a flow control valve.

Fluid addressing system 120 is illustrated schematically in FIG. 4 .Fluid addressing system 120 can route the output solution from thefluidic synthesizer 110 to a target location on microphysiologicalplatform 100. For example, fluid addressing system 110 can divert theoutput solution to at least one microphysiological device 130. As shownin FIG. 4 , fluid addressing system 120 can function as a demultiplexercomponent, to selectively route fluid (e.g. the output of fluidsynthesizer 110) received at fluid input 122 to any one or more ofmultiple fluid outputs 124, under the control of control or select lines125. The control lines 125 include control line 1, up to control line S.The control lines 125 control the valves 126A, 126B, 126D, 126E, 126F(collectively referred to as valves 126) to be open or closed. As shown,the valves 126 are arranged in pairs. As an example, the control lines125 can include “high” lines and “low” lines, where the high linescontrol the first of each pan of valves 126 and the low lines controlthe second of each pair of valves 126. For example control line 1 caninclude a high line and a low line. The high line of control line 1 cancontrol valve 126A while the low line of control line 1 can controlvalve 126B. Control line S can include a high line that controls valve126C and valve 126E as well as a low line that controls line 126D andline 126F. An advantage of such a system is the ability to controlmultiple valves with a single line. However, the open/closed status ofsome of the valves 126 would be coupled together. In other words, itwould not be possible to control each of the valves 126 independently.For example valve 126C and valve 126E would have the same open/closedstatus, while valve 126D and valve 126F would have the same open/closedstatus. In some embodiments, one or more of the valves 126 can be adome-shaped membrane valve connected to the at least onemicrophysiological device 130 through a rectangular-profilemicrochannel. In certain embodiments, one or more of the valves 126 canoperated by positive pressure. In certain embodiments, one or more ofthe valves 126 can be operated by the application of a vacuum. Incertain embodiments, one or more of the valves 120 can be operated by acombination of positive and negative pressure. In certain embodimentsone or more of the valves 126 can utilize rectangular profilemicrochannels.

In some embodiments, the fluid addressing system 120 may be implementedwith a pneumatically gated or valved demultiplexer component to selectoutput channels (fluid outputs 124). The demultiplexer can operate usingbinary addressing logic with integrated flow valves. For example, a “0”can symbolize a closed valve, and a “1” can symbolize an open valve.Each downstream microphysiological device 130 (or other devices) in themicrophysiological platform can have a numeric address whose binaryrepresentation can encode the valve states required to open the fluidaddressing system output that corresponds to it. The binary addressinglogic can be controlled by a plurality of pneumatic or hydraulic controlor select lines 125. The mathematical relationship between the quantityof select lines (S) and the quantity of addresses (A) can be describedby A=2^(S). In some embodiments, two pneumatic or hydraulic connectionscan be used to control each select line, in which case the mathematicalrelationship between the quantity of select lines (S) and the quantityof addresses (A) can be described by A=2^(S+1). For example, in animplementation in which the binary addressing logic has six select linesthere are 64 potential output addresses. Of course, the binaryaddressing logic can have fewer than six select lines, or it can havemore than six select lines (e.g., 10 select lines to handle 1024addresses). In some embodiments, each select line can be operated by anexternal 4-way valve, by two external 3-way valves or two external 2-wayvalves. In non-limiting embodiments, the binary addressing system caninclude 7 total 2-way, 4-port solenoid valves as pilot valves (e.g.,2⁷=128 individually addressable fluid outputs or outputs from the fluidaddressing system), or 14 total 2-way, 4-port solenoid valves as pilotvalves (e.g., 2¹⁴=16,384 individually addressable fluid outputs oroutputs from the fluid addressing system).

In some embodiments, the fluid addressing system can contain a “flush”output 127 to wash any liquid or dead volume currently contained in thefluid addressing system or downstream channels. In some embodiments, theflush output 127 can be fluidically coupled to the effluent channel 160,as described above with reference to FIG. 1 . In some embodiments, theflush output 127 can be fluidically coupled to the fluid outputs 124.Fluid addressing system 120 provides great flexibility for operation ofthe microphysiological platform 100 and delivery of customized fluidand/or wash fluid to one, more than one, or all devices (such asmicrophysiological devices 130) downstream of fluid addressing system120. For example, in some embodiments, fluid addressing system 120 caninclude an “off” state in which all outputs 124 are blocked. Fluidaddressing system 120 can also open one of its fluid outputs 124, asubset of more than one of its fluid outputs 124, or all of its fluidoutputs 124. In some embodiments, when the fluid addressing system 120is delivering a fluid from its fluid input 122, the fluid addressingsystem 120 can simultaneously open all or a subset of the outputchannels connected to microphysiological devices requiring said fluid,and in doing so perfuse all of the selected microphysiological devicessimultaneously.

Accordingly, the customized fluid mixture by fluidic synthesizer 110 canbe delivered to one or a plurality of microphysiological devices 130 bythe fluid addressing system 120. Fluid addressing system 120 can openflow valves to the selected microphysiological device or devices 130,allowing the customized fluid mixture produced in the fluidicsynthesizer 110 to flow into a target microphysiological device ordevices 130 while preventing it from entering any othermicrophysiological device or devices 130.

In some embodiments, fluid addressing system 120 can include a bypassselector 128 that can control the flushing of all fluid upstream of aselected microphysiological device 130. For example, downstream of themost downstream select or control line 125 in the fluid addressingsystem 120, the fluid can continue directly to a targetmicrophysiological device 130 or can be bypassed by the bypass selector128 into a waste stream without disrupting the target microphysiologicaldevice 130. In other words, the bypass selector 128 can determinewhether fluid output from the fluid addressing system 120 reaches itstarget location in the array of microphysiological devices 130 or if itswitches to a waste stream and is flushed out. This reduces the amountof waste in the event that the user does not want to deliver the fluidcurrently in the fluid addressing system 120 into the microphysiologicaldevices 130. With the bypass selector 128 only the fluid in the fluidaddressing system 120 and upstream thereof would need to be flushed andgo to an effluent or waste stream, rather than also including the fluidin the microphysiological devices 130 as well. The binary addressinglogic can be used to minimize the requirement for solenoid valves tocreate a microphysiological platform 100 for high-throughput screeningor testing on a multitude (e.g., hundreds to tens of thousands) ofmicrophysiological devices 130. The microphysiological platform 100 canbe an organ-on-a-chip system that can include at least oneorgan-on-a-chip.

In some embodiments, the fluid addressing system 120 can route the flowof a fluid to a different layer of microphysiological platform 100. Insome embodiments, multiple layers of microphysiological platform 100 canalternate between rounded-profile or domed-profile microfluidicchannels, which can be fabricated by photoresist-reflow molding, andsquare-profile or rectangular-profile microfluidic channels, which canbe fabricated by negative-photoresist molding. Each of therounded-profile, domed-profile, square-profile, and rectangular profilemicrofluidic channels can be located on the same layer or on differentlayers of microphysiological platform 100. Such alternating between themicrofluidic channels can permit compatibility between membrane valvesoperating in rounded-profile microfluidic channels andmicrophysiological devices 130 operating in rectangular-profilemicrofluidic channels. In some embodiments, microphysiological platform100 can include rectangular-profile microfluidic channels used inconjunction with dome-shaped valves. In some embodiments, therectangular fluid lines or microfluidic channels can be decoupled fromany swelling of pneumatically driven select lines or hydraulicallydriven select lines. The swelling can subject the pneumatic or hydraulicselect lines to cross-sectional distortion when higher pressure is usedto enable more complete and/or rapid membrane valve actuation. In someembodiments, a fluid stream can be transferred from a first flow layerto a second discrete valving layer (“out-of-plane” or “off-layer”valving) and then returned to the first flow layer within the fluidaddressing system. The out-of-plane or off-layer valving can allowpneumatic or hydraulic control valves to be placed anywhere on thefootprint of microphysiological platform 100. For example, the pneumaticor hydraulic control valves can be placed without unintendedinterference or pressurized deflection of fluid flow layers at locationsother than those intended for valving purposes.

In some embodiments, fluids can flow between layers through holes (e.g.,“vias”), which can be contiguous between the microfluidic channelspresent in opposing layers of the device. The placement of vias can beutilized to allow a first fluid channel to cross at least one secondfluid channel without fluid contact by routing the first fluid channelon a first layer vertically to a second layer, then running the firstfluid channel laterally across the second fluid channel and thenvertically back to the first layer to create an overpass or underpass.In some embodiments, the vias can be utilized to transport fluid betweena microchannel with a rectangular cross-section to a microchannel with arounded cross-section. Pneumatically hydraulically controlled membranevalves can be placed within the layers with rounded cross-sections, suchthat the actuated membrane can form a more robust seal along the roundedcontour of the valved microchannel.

As described above with reference to FIG. 1 , microphysiologicalplatform 100 can include one or more arrays of microphysiologicaldevices 130. Each such microphysiological device 130 can containmicroengineered environments designed to stimulate physiologicallyrelevant tissue or organotypic development. In some embodiments, all ofthe microphysiological devices 130 in the array, and/or on the entiremicrophysiological platform, can share the same generic design. In otherembodiments, microphysiological devices 130 can have multiple designsand be distributed over different positions in an array, or in multiplearrays on microphysiological platform 100.

Each microphysiological device 130 in microphysiological platform 100can be configured to contain, and can include, any one or more oftissues, cells, bacteria, viruses, other living entities, biologicalscaffolds, or explanted tissues, and can replicate the structure andfunction of an organ, or part thereof. In some embodiments, one or moreof the microphysiological devices 130 can start as a bare structure.Thus, in an initial state of a microphysiological platform 100, it maybe devoid of any biologic material, and microphysiological devices 130are configured to receive such material, and then to culture or grow it.Therefore, in other states of a microphysiological platform 100, it maycontain biologic material in one or more microphysiological devices 130,in any one or more stages of physiological development. Thus, in someembodiments, one or more of the microphysiological devices 130 can beseeded with tissue by introducing cells that can be fed culture mediumto grow into tissue or organs. Such cells can be introduced into themicrophysiological device by any one or more of several routes. In someembodiments, cells can be introduced directly into themicrophysiological device, e.g. by fluid handler 195. Alternatively, orin addition, cells can be introduced into any one or more other portionsof microphysiological platform 100 that are fluidically coupled (orcoupleable) to a microphysiological device 130, such as into, orupstream of, the fluidic synthesizer 110, into, or upstream of, thefluid addressing system 120, etc.

In some embodiments, the substrate 101 can include functionalized sites.In some embodiments functionalized sites on the substrate 101 can befunctionalized such that the cells can grow more easily. In someembodiments, the functionalized sites can be formed on the substrate 101before the substrate 101 is enclosed to form the microphysiologicaldevice 130. In some embodiments, the functionalized sites can be formedin the microphysiological device 130 while the substrate 101 is insidean enclosure or “hive”. In some embodiments, functionalized sites on thesubstrate 101 can be formed in a monolithic structure. In someembodiments, functionalized sites on the substrate 101 can bedisaggregated across multiple components or structures.

In some embodiments, the cells can include heart bone cells, kidneycells, liver cells, gut cells, lung cells, or any combination thereof.In some embodiments, the microphysiological devices 130 can bestructured to simulate heart tissue, bone tissue, kidney tissue, livertissue, gut tissue, lung tissue, or any combination thereof.

FIGS. 5A-5C are schematic illustrations of an embodiment of amicrophysiological device 130 that may be included in themicrophysiological platform 100 of FIG. 1 . As shown, themicrophysiological device 130 includes tissue chambers 137 interposedwith channels 136. Each tissue chamber 137 can, in some states ofmicrophysiological device 130 and microphysiological platform 100,contain cells, organ tissue cultures tissue, etc. (and thus tissuechamber 137 can also be referred to as tissue 137, as appropriate in thecontext). In some embodiments, the tissue chambers 137 can be in fluidiccommunication with the channels 136. In some embodiments, the tissuechambers 137 can include fluidic pathways, in which tissue cultures cangrow. In some embodiments, fluid and/or nutrients can flow freelybetween the tissue chambers 137 and the channels 136. In someembodiments, the channels 136 can be used to deliver fluid to the tissuechambers 137 and/or tissue contained therein. In some embodiments,membranes can be placed between the tissue chambers 137 and the channels136, or can be placed between two or more different tissue types withina tissue chamber 137. In some embodiments, the membranes can includesemi-permeable membranes. In some embodiments, fluids in the tissues 137and the channels 136 can be held in place by surface tension rather thanmembranes. In other words, there can be open fluidic communicationbetween the channels 136 and the tissues 137.

As shown, the tissue chambers or tissues 137 include tissue chamber ortissue 1 up tissue chamber or tissue N, while the channels 136 includechannel 1 up to channel N, where N is an integer. In some embodiments, Ncan be 1, 2, 3, or more, up to any practical limit, and to meet therequirement for any desired organ-on-a-chip or other structure. In someembodiments, the microphysiological device 130 can include N tissues 137and N−1 channels. In other words, the tissues 137 can be on the outeredges of the array of tissues 137 and channels 136 such that the thereis one less channel 136 than tissues 137. In some embodiments, themicrophysiological device 130 can include N tissues 137 and N+1 channels136. In other words, the channels 136 can be on the outer edges of thearray of tissues 137 such that there is one less tissue 137 than channel136.

As shown, tissue 1 includes tissue 1 fluid input A (132-1A) from thefluid addressing system 120 and optional tissue 1 fluid input B (132-1B)from the fluid handler 195, channel 1 includes channel 1 fluid input A(133-1A) from the fluid addressing system 120 and optional channel 1fluid input B (133-1B) from the fluid handler 195, tissue N includestissue N fluid input A (132-NA) from the fluid addressing system 120 andoptional tissue N fluid input B (132-NB) from the fluid handler 195, andchannel N includes channel N fluid input A (133-NA) from the addressingsystem 120 and optional channel N fluid input (133-NB) from channel Nfluid input B. In some embodiments, the inputs (132-1A, 132-1B, 133-1A,133-1B, 132-NA, 132-NB, 133-NA, 133-NBA collectivity referred to hereinafter as inputs 132,133) can include injection ports In someembodiments, the inputs 132, 133 can include valves to regulate fluidflow.

As shown, tissue 1 includes tissue 1 fluid output 138-1, channel 1includes channel 1 fluid output 139-1, tissue N includes tissue N fluidoutput 138-N, and channel N includes channel N fluid output 139-N. Asshown each of the outputs (138-1, 139-1, 138-N, and 139-N, collectivelyreferred to herein as outputs 138, 139), are configured to discharge anoutput solution, the composition of which is based on the composition ofthe fluid solution received from the fluid output of the fluidicsynthesizer, via the fluid addressing system, at the respective fluidinput to the channel, as may be affected, modified and/or supplementedby interaction of the fluid solution with any tissue in the respectivetissue chamber that is in fluidic communication with the respectivechannel. In some embodiments, the outputs 138, 139 can recycle thefluids exiting the tissues 137 and the channels 136. In someembodiments, the outputs 138, 139 can feed to waste streams. In someembodiments, the outputs 138, 139 can feed to further processing units.As described in more detail below, each tissue chamber 137 or channel136 can include, or be fluidically coupled to, a separate sensor or setof sensors, FIG. 5B shows the tissue 137 and channels 136 oriented in avertical configuration. In other words, the channels 136 are positionedabove and below the tissue 137. FIG. 5B includes axes, where the x-axisis horizontal and the z-axis is vertical. In FIG. 5B, fluid would flowhorizontally through the tissue 137 and the channels 136. FIG. 5C showsthe tissue 137 and the channels 136 oriented in a horizontalconfiguration FIG. 5C includes axes, where the x-axis is horizontal andthe z-axis is vertical. In FIG. 5C, fluid would flow vertically throughthe tissue 137 and the channels 136.

Each or any of the microphysiological devices 130 can be implemented asany of a non-limiting variety of organ-on-a-chip models, including aneye model (e.g., the model(s) disclosed in U.S. Pat. No. 10,360,819), aplacenta model (e.g., the model(s) disclosed in U.S. Pat. No.10,633,623), a lung model (e.g., the model(s) disclosed in U.S. Pat.Pub. Number 2018/0216058), an organ fibrosis model (e.g., the model(s)disclosed. in U.S. Pat. Pub. Number 2018/0230415), a cervix model (e.g.,the model(s) disclosed in U.S. Pat. Pub. Number 2018-0312810), avasculature model (e.g., the model(s) disclosed in WIPO Pub. Number2019/191111), and/or a pulmonary airway model (e.g., the model(s)disclosed in WIPO Pub. Number 2020/073043). Further, each or any of themicrophysiological devices 130 can also include certain non-limitingfeatures for organ-or-a-chip models, including a decellularizedextracellular matrix (e.g., as disclosed in U.S. Pat. Pub. Number2018/0126037), a heterobifunctional crosslinker (e.g., as disclosed inU.S. Pat. Pub. Number 2018/0223251) and/or a native extracellularmatrix-derived membrane insert (e.g., as disclosed in U.S. Pat. Pub.Number 2020/0190456). The disclosure of each of the foregoing patentsand published patent applications is incorporated herein by reference.

As described above with reference to FIG. 2 , observations of themicrophysiological devices 130, e.g. of the biological material, tissue,tissue structures, etc. contained therein, can be acquired by one ormore imagers disposed in, associated with, and/or controlled by thecontrol system. Imaging modalities or methods that may be implemented bysuch imagers can include optical microscopy, magnetic resonance imagingor methods of computed tomography (CT) scanning. Correspondingly, thestructure of each microphysiological device 130 from which image data isto be acquired should be configured to permit effective acquisition ofimages by the desired imaging modality. For example, optical microscopyrequires that the structure of the microphysiological device 130 and,any other structure of microphysiological platform 100 between theimager and the microphysiological device 130 be sufficientlytransmissive of the light frequencies to be employed by the opticalmicroscope, and that indices of refraction any different materials, andincident angles of light at interfaces thereof, enable adequate imaging.The data acquired from the use of one or a combination of said imagingmethods on the contents of a microphysiological device 130 can be usedfor nonlimiting analytical purposes including one or a combination ofthe following: creating three-dimensional reconstructions of thebiological entities in the microphysiological devices; applyingcomputational measurement techniques to the acquired data to obtainphenotypic data about the biological specimen or entities; measuring thedeviation of observed phenotype or physiology from the expectedphenotype or physiology; or measuring dynamic behaviors of the observedbiological entities over time, including, for purposes of illustrationand not limitation, the rate of cell division, the formation ofbiological networks, or the development and function of biologicaltissues, organs, or functional subunits of organs.

Other sensor types can include an electronic sensor that monitorssecondary or derived data (e.g., software reconstructions or models ofbiological or physical entities), or combinations thereof.

In addition to information gathered from sensors, such as the imagersdescribed above, that are not part of the microphysiological platform100, information about the contents of each microphysiological device130 can also be gathered with one or more sensors coupled to orincorporated into microphysiological platform 100. Such sensors can beincorporated in whole or in part, in the microphysiological device 130and/or can be incorporated, in whole or in part, in one or morebiosensors 140 that may be operatively, e.g. fluidically, coupled(continuously or selectively) with the fluid output 134 of themicrophysiological device 130. The sensor(s) can be placed locally todownstream of, or upstream of each microphysiological device 130.

In same embodiments, microphysiological device 130 can include at leastone integrated sensor, which can include a chemical sensor, a mechanicalsensor, an optical sensor, or combinations thereof. In some embodiments,the integrated sensor can be incorporated in-line with one or more ofthe microphysiological devices 130. The at least one sensor cantransduce one or a combination of signals. The signals can be originatedfrom the microphysiological device, the periphery of themicrophysiological device, an experimental factor or conditionassociated with the microphysiological device/platform, or combinationsthereof.

In some embodiments in which the sensor is a chemical sensor, whetherintegrated with of placed downstream of microphysiological device 130,the sensor or sensors can transduce chemical signals. Such chemicalsignals can include the presence and concentration of biologicalsecretions (e.g., hormones, paracrine factors, extracellular matrixcomponents, metabolic byproducts), a biological consumption or uptake ormodification of a chemical substrate (e.g., a consumption, uptake, ofmodification of nutrients, glucose, amino acids, biologicaltherapeutics, drugs, drug products, toxins, gases, dissolved solids,dissolved chemical compounds, or aerosolized compounds), a fluid's pH, afluid's osmolarity, a fluid's chemical composition (name andconcentration of chemical constituents), multiplexed chemical sensing ofa plurality of chemical targets (e.g., tens or hundreds of proteins bycompetitive binding against a dotted antibody microarray), environmentalgas compositions, or combinations thereof. In non-limiting embodiments,the biological consumption or uptake or modification of a chemicalsubstrate can be determined by measuring a downstream concentration offluid effluent from a microphysiological device 130 against aconcentration in fluid upstream of microphysiological device 130.

In some embodiments in which the sensor is a mechanical sensor, whetherintegrated with or placed downstream of microphysiological device 130,the sensor or sensors can transduce mechanical signals including anexertion of mechanical force or forces by microphysiological device 130(e.g., a contraction of muscle tissue or pressure imposed by a modeledtumor's growth), application of mechanical force or forces ontomicrophysiological device 130 (e.g., closed-loop pressure control orclosed-loop electromechanical actuation), ambient pressure, materialproperties (e.g., a modulus of elasticity, a shear modulus, and cyclicloading characteristics), or combinations thereof.

In some embodiments in which the sensor is an optical sensor, whetherintegrated with or placed downstream of microphysiological device 130,the sensor or sensors can transduce optical signals including imagesacquired by methods of light transmission microscopy, images acquired byfluorescent microscopy (e.g., confocal microscopy, light-sheetmicroscopy and super-resolution microscopy), quantitative opticalmeasurements (e.g., transmittance, luminescence,electrochemiluminescence fluorescence, and fluorescence resonance energytransfer (FRET) techniques), phenotypic observations, observations ofmorphology and physiology in 3D computer reconstructions orreprojections, images of structural motifs (e.g., scaffold-taggedfluorescent markers, embedded quantum dots, and second harmonicgeneration (SHG) imaging) or combinations thereof.

In some embodiments, microphysiological device 130 can contain acombination of chemical sensors, mechanical sensors, optical sensors,and modality-specific sensing methods. In non-limiting embodiments,additional sensor modalities can be integrated or applied. Theadditional sensor modalities can include magnetic resonance, X-raytransmission, computed tomography, electronic monitoring of a softwarereconstruction, software reconstructions of a microphysiological device,combinations thereof. In some embodiments, a microphysiological devicecan include one or a plurality of surface plasmon resonance (SPR)biosensors against one or a plurality of corresponding target analytes,alone or in conjunction with one or a combination of the sensorspreviously listed.

When any fluids can be delivered to microphysiological device 130, anequal volume of the existing fluids can be displaced over the sensoritself, over the area to which a sensor is sensitive, or into the volumeto which a sensor is sensitive, which can lie downstream ofmicrophysiological device 130, e.g. part of a biosensor 140.

As shown schematically in FIG. 1 , microphysiological platform 100 caninclude an array of biosensors 140, and the biosensors 140 can befluidically coupled to microphysiological devices 130. The fluidiccoupling arrangement can be implemented in different ways. For example amicrophysiological device 130 can have a single biosensor 140 with adedicated fluidic coupling therebetween, i.e. the biosensor 140 can onlysense properties of fluid output from the device 130. In someimplementations the relationship between microphysiological devices 130and biosensor 140 can be one-to-many (multiple biosensors can befluidically coupled, or coupleable, to a single microfluidic device130), many-to-one (a single biosensor can be fluidically coupled, orcoupleable, to multiple microfluidic devices 130), or many-to-many,using appropriate microfluidic couple, switching, multiplexing,demultiplexing, etc. arrangements, as represented schematically in FIG.1 in the fluid paths between the array of microphysiological devices 130and he array of biosensors 140. Any or all of these relationshipsbetween microphysiological devices 130 and biosensors 140 can be used ona single microphysiological platform 100. Correspondingly,microphysiological platform 100 can include multiple types of biosensors140. For example, microphysiological platform 100 can includemultiplexed biosensor which can be electrochemical biosensors, opticalbiosensors, and/or fluorescence biosensors. The types of biosensors 140patterned adjacent to microphysiological devices 130 inmicrophysiological platform 100 can differ. For example, a firstmicrophysiological device 130 can be coupled with an opticalfluorescence-based biosensor 140, a second microphysiological device 130can be coupled with a pH sensor 140, and third microphysiological device130 can be coupled with a trans-epithelial electrical resistance (TEER)sensor 140. In some embodiments, a first microphysiological device 130can have a first combination of sensor modalities that is different froma second combination of sensor modalities associated with a secondmicrophysiological device 130. For example, the variability incombinations of sensor modalities or variability in sensor targets canexist between microphysiological devices 130 that are contained withinthe same microphysiological platform 100.

In some embodiments, a biosensor 140 can be implemented as, for example,a label-free microfluidic biosensor, and can be either co-localized withor located downstream of a microphysiological device 130. In someembodiments, a continuous acquisition from a plasmonic biosensormultiplexed for multiple biological target analytes can allow thereal-time characterization of the microenvironment and secretome of themicrophysiological device 130 with high temporal resolution and avoidthe cost of label-based biosensors, whose cost of operation (primarilythe consumption of labeled molecules) scales with operational duration.One example of such an implementation is microphysiological device 230and biosensor 240 shown in FIGS. 6A to 6D. In this embodiment, a surfaceplasmon resonance (SPR)-based biosensor 240 is disposed in closeproximity to microphysiological device 230 for continuous monitoring oftarget analytes in the fluid flowing through the organ-chip environment.In some embodiment, the biosensor 240 can be placed downstream of themicrophysiological device. In some embodiments, the biosensor 240 canmake measurements in-line with the microphysiological device 230. Insome embodiments, the biosensor 240 can be integrated into themicrophysiological device 230. In some embodiments, the biosensor 240can include a colorimetric based sensor. In some embodiments, thebiosensor 240 can include a fluorescence based sensor. In someembodiments, the biosensor 240 can be transduced by a by a microscope.In some embodiments, the biosensor 240 can be electric (e.g., cyclicvoltammetry). In some embodiments, the biosensor 240 can be transducedby a potentiostat. In some embodiments, the microphysiological device230 can include two inlets integrated into the microphysiological device230. As described above with reference to FIG. 5B and FIG. 5C, theinlets can be oriented on top and on bottom of the microphysiologicaldevice 230. In some embodiments, the inlets can be oriented on eitherside of the microphysiological device 230. In some embodiments, amultiplexed biosensor 140 can include a thin film and biorecognitionmolecules. The thin film can be deposited on the biosensor 140 through aphysical vapor deposition technique. For example, physical vapordeposition of a gold thin-film can be used to generate the resonantplasmonic coupling with laser emission at a specific, tightly controlledwavelength and critical incident angle. The surface can be spotted withbiorecognition molecules (e.g., antibodies, DNA, RNA, XNA, etc.) thatcan be selective to the biomolecules of interest in a solution. Bindingof the molecules-of-interest to the biorecognition elements adsorbed tothe gold surface can produce surface loading, the extent of whichcorresponds to a shift in the critical angle at which the resonantcoupling can occur. By tracking this angle for each of the spots or byusing a camera to image them all at once, the concentration of eachanalyte of interest in the solution can be measured without anyadditional labels being required. The biosensor 140 can be a label-freebiosensor that can avoid the detrimental effects of the possiblelabeling-molecule toxicity to tissue culture. In some embodiments,biosensor 140 can include multiple biorecognition elements that candetect more than one target analyte at the same time.

In certain embodiments, the biosensor 140 can be integrated into atleast one microphysiological device 130 to permit on-chip, real-time,and high content data acquisition with reduced sample volumerequirements. The biosensor 140 can automatically sample a fluid contentin microphysiological devices 130 and quantify target analytes inreal-time. Similar to a barcode reader, for example, the biosensor 140and an optical transducer can sample and quantify a fluid content usinga laser beam reflection as it scans across the backside of the SPRspots. Accordingly, the biosensors 130 with the SPR can have moreunproved data-capture dimensionality than ELISA and microscopy analyses.

In certain embodiments, biochemical,electrochemical/electrochemiluminescent, mechanical, or/and opticalbiosensing schemes can be incorporated into microphysiological platform100 through the modification of the components of biosensor 140 or theiron-chip placement. For example, given that the multiplexed biosensorzones can utilize a patterned gold deposition on glass, variousbiosensing schemes can be incorporated into the same gold pattern layerwithout additional fabrication processes. For example, a series ofinterdigitated microelectrode arrays for redox coupling-basedelectrochemical biosensing can be achieved through the disclosedtechnique.

In some embodiments, the measurements or readouts produced by biosensor140 can be used for chemical or biological characterization of thebiological entities being cultured or experimented within amicrophysiological device 130. This chemical or biologicalcharacterization can be based the appearance, disappearance, or changeof a concentration of one or more compounds. For example, the compoundcan include chemical or biological compounds, chemical or biologicalmoieties, dissolved compounds, biologically secreted compounds,metabolized or altered compounds, target analytes, or combinationsthereof. In some embodiments, this characterization can be made as acomparison between the composition of a solution at a first locationupstream of a microphysiological device 130 and the composition of thesolution at a second location downstream of the microphysiologicaldevice 130. In some embodiments, this measurement can be used to infer,measure, or deduce one or a combination of characterizing observationsor factors about the microphysiological device 130. For example, theobservations can include (1) the uptake of compounds, chemicals, drugs,or biological agents from the solution into the biological entities in amicrophysiological device 130; (2) the uptake of complex biologicalentities, including extracellular vesicles, exosomes, viral particles,constituents an extracellular matrix, polymers, enzymes, nucleic acids,peptides, or proteins into the biological entities in amicrophysiological device 130, (3) the secretion of compounds,chemicals, or biological agents from the biological entities in amicrophysiological device 130 into the surrounding fluid solution; (4)the secretion of complex biological entities, including extracellularvesicles, exosomes, viral particles, constituents of an extracellularmatrix, polymers, enzymes, nucleic acids, peptides, proteins from thebiological entities a microphysiological device into the surroundingfluid solution, (5) the modification of compounds, chemicals, drugs, orbiological agents in the solution by the biological entities in amicrophysiological device 130; or (6) the modification of complexbiological entities, including extracellular vesicles, exosomes, viralparticles, constituents of an extracellular matrix, polymers, enzymes,nucleic acids, peptides, or proteins by the biological entities in amicrophysiological device 130.

As shown schematically in FIGS. 1 and 2A-2B, in some embodiments, one ormore sensor data transmitters 150 can be associated with one or more ofthe biosensors 140, to enable a treasured datum or data from biosensor140 to transmitted to a device external to microphysiological platform100, such as data receiver 181 that is part of a control system 170.This is also illustrated schematically in FIGS. 7A and 7B. FIG. 7Aillustrates an exemplary diagram of the augmentation of discretemicrophysiological devices 330, each integrated with an on-chip fluidicsynthesizer 310. The microphysiological devices 330 further includechemical sensors or biosensors 340 located downstream of themicrophysiological device 330. Digitally transduced data from one or aplurality of the sensors can be transmitted by sensor data transmitter350 to an electronic device, such as computer C for continuous automaticmonitoring, and data processing. FIG. 7B shows an exemplary diagram ofthe integration of multiple discrete microphysiological devices 430 intoa single monolithic microphysiological platform 400, in winch the fluidoutput from a single fluidic synthesizer 410 can be addressed to one ofa plurality of selected individual microphysiological devices 430 usinga fluid addressing system 420. Downstream of the individualmicrophysiological devices 430 lie biosensors 440, which can allowtransduction of the fluids partial or whole chemical composition andsubsequent transmission of said compositional measurement ormeasurements by sensor data transmitters 450 to an electronic devicesuch as computer C for data monitoring, data processing, or datamonitoring and processing.

Returning to FIGS. 1 and 2A-2B, in some embodiments, a biosensor 140with a sensor data transmitter 150 can characterize the concentrationsof a mixture of fluid constituents at certain given positions andtransmit this information electronically to a fluidic synthesizer, suchas fluidic synthesizer 110, at a different position. This differentposition can refer to a different position the same microphysiologicalplatform 100, to another microphysiological platform 100 engaged withthe same control system, or to another microphysiological platform orother device altogether. Then, the fluidic synthesizer can dynamicallyresynthesize this same fluid mixture and outflow it. Since theconnection can be electronic, the link between the sensor and thefluidic synthesizer can span just several millimeters on the samephysical device (e.g. on the same microphysiological platform 100), orthe link can span two or more of the disclosed microphysiologicalplatforms located across the world from one another. This enablesfunctionality described in more detail below.

The microphysiological platform 100 described above provides thecapability for the fluidic synthesizer 110 to create a first fluidmixture be delivered by fluid addressing system 120 to a firstmicrophysiological device 130. Then, the fluidic synthesizer 110 cancreate a second fluid mixture that can subsequently be delivered byfluid addressing system 120 to a second microphysiological device 130.By this operation, microphysiological platform 100 can sequentiallydeliver at least one unique fluid mixtures to at least onemicrophysiological device 130 integrated into the microphysiologicalplatform 100. In some embodiments, this operation can be cycledautonomously (e.g. under the control a a main controller of controlsystem) to create an automated microphysiological platform 100. In someembodiments, fluidic synthesizer 110 can produce multiple fluid mixturessequentially, and fluid addressing system 120 can route the mixtures tothe designated microphysiological devices 130. In some embodiments, eachmicrophysiological device 130 can have its own fluidic synthesizerdelivering a unique fluidic mixture. Alternatively, multiple fluidicsynthesizers 110 incorporated into microphysiological platform 100 canoperate simultaneously at multiple positions.

In some embodiments, microphysiological platform 100 can performdiagnostic procedures to attempt to detect a component failure, animpending component failure, or a component anomaly. Such diagnosticprocedures can be performed at various time points. For example, adiagnostic procedure can be performed preemptively prior to the start ofan experiment, at one or a plurality of discrete intervals during thecourse of an experiment, continuously during the course of anexperiment, at the conclusion of an experiment, or a combinationthereof. In some embodiments, such diagnostic procedures can include thecharacterization fluids through components (e.g., microphysiologicaldevices 130) in the microphysiological platform 100, the actuation ofvalves or flow controls in the microphysiological platform 100, themeasurement of one or a plurality of sensor readings, the measurement offluid flow rates, the measurement of electrical properties, automated orsemi-automated optical inspection, or combinations thereof. In someembodiments, microphysiological platform 100 can passively monitorheuristics, performance measurements, performance metrics, or acombination thereof against expected or nominal values to monitor itselffor component failures or component anomalies without activelyperforming diagnostic procedures.

One exemplary implementation of microphysiological platform 100 is shownin FIGS. 8A to 8D. FIG. 8A shows a plan view of a microphysiologicalplatform 500. As shown in FIG. 8A, microphysiological platform 500 isimplemented on a flat, circular substrate 501 with a peripheral edge504. As shown in FIG. 8A, in this exemplary embodiment, the components amicrophysiological platform 500 have a tiled or mirrored layout (whichcan be analogous to certain tiled instruction-executing cores of amulti-core microprocessor). That is, each half microphysiologicalplatform includes a fluidic synthesizer 510, a fluid addressing system520, an array of microphysiological devices 530, and an array ofbiosensors 540. The two halves of the microphysiological platform sharea common effluent channel 580.

In this embodiment, each microphysiological device 530 is paired in aone-to-one relationship with a dedicated biosensor 540 (similar to theembodiment shown in FIGS. 6A to 6D. Each array of microphysiologicaldevices 530 includes 64 devices, and correspondingly each array ofbiosensors 540 includes 64 biosensors. FIG. 8B shows a close-up view ofthe fluidic synthesizer 510. As shown, the fluidic synthesizer 510includes fluid inlet ports 512, valve control ports 517, and mixingchamber. As shown, there is a 1:1 relationship between the fluid inletports 512 and the number of fluidic lines that go into the mixingchamber 516. FIG. 8C shows a close-up view of portions of the fluidaddressing system 520, as well as portions of the array ofmicrophysiological devices 530 and portions of the array of biosensors540. As shown, the fluid addressing system 520 includes fluid inputs522, fluid outputs 524, control lines 525, and valves 526. The fluidinlet ports 312 feed to the fluid inputs 522, while the valve controlports 517 and the control lines 525 control the valves. In someembodiments, the microphysiological devices 530 and the biosensors 540can be the same or substantially similar to the microphysiologicaldevice 230 and the biosensors 240, as described above with reference toFIGS. 6A-6D. As shown, the biosensors 540 can be connected to themicrophysiological devices 530 via a serpentine flow path.

FIG. 8D shows a close-up view of an interface between the fluidaddressing system 520 and the array of microphysiological devices 530.As shown, the microphysiological devices 530 include tissue chambers ortissues 537, channels 536, tissue input 532A, tissue input 532B, andchannel input 533B. A fluid handler 595 is also shown. In someembodiments, the tissue input 532A can be the same or substantiallysimilar to inputs 132-1A or 132-NA, as described above with reference toFIG. 5A. In other words, fluids can pass between the fluid addressingsystem 520 and the tissue chamber of tissue 537 via the tissue input532A. In some embodiments, the tissue input 532B can be the same ofsubstantially similar to inputs 132-1B of 132-NB, as described abovewith reference to FIG. 5A. In other words, fluids can pass between thefluid handler 595 and the tissue 537 via the tissue input 332B. In someembodiments, the channel input 5338 can be the same or substantiallysimilar to inputs 133-1B or inputs 133-NB, as described above withreference to FIG. 5A. In other words, fluids can pass between the fluidhandler 595 and the channels 536 via the channel input 533B. In someembodiments, the microphysiological devices 530 can include inlets thatcreate fluidic paths between the fluid addressing system 520 and thechannels 536 (e.g., the same or substantially similar to the inputs133-1A and 133-NA, as described above with reference to FIG. 5A). Anycombination of the aforementioned fluidic couplings can be included inthe microphysiological platform 500.

In some embodiments, such as that shown in FIGS. 8A to 8D, the twoinstances of the components microphysiological devices 530 and/orsensors 540 associated with microphysiological devices 530) can beidentical. This layout can allow one instance to function independentlyof one or a plurality of other instances for increasing operationalthroughput or operating in an identical-copy configuration forredundancy. For example, while biological replicates can be added tomicrophysiological devices 530 to improve statistical rigor or toincrease reproducibility, mirroring several components of the entiresystem across both sides to create two instances of “cores” can permitfault tolerance for operational anomalies (e.g., inlet clogging, fluidleakage, valve puncture, component failure, or combinations thereof). Insome embodiments, the mirrored or tiled components can be disabledwithout affecting other patterned instances. This redundancy can allowdevices with manufacturing defects to be “binned” or operationallyrestricted to a lower capability or feature set. Such binned orrestricted microphysiological platforms can be used for variouspurposes. For example, the binned or restricted microphysiologicalplatforms can be sold or marketed at different tiers of technologicalcapability. Such binned or restricted microphysiological platforms canoperate or complete portions of an ongoing experiment following thepartial failure of one or a plurality of component. Such binned orrestricted microphysiological platforms can also serve as a substratefor a test, calibration, demonstration, or sampling procedures.

Although the microphysiological platform 500 shown in FIGS. 8A. to 8Dincludes two instances of identical sets of components, in otherembodiments more than two instances of components can be included, e.g.three, four, or more.

Fluid “Teleportation”

As noted above, the microphysiological platform architecture describedherein provides for powerful capabilities. One important capability is“fluidic teleportation,” “digital teleportation,” or virtual fluidiccoupling between components that are on the same microphysiologicalplatform or distributed across multiple platforms that arephysically/geographically separated (e.g. multiple microphysiologicalplatforms engaged with the same control system, engaged with differentcontrol systems in the same room, facility, etc., of engaged withcontrol systems spaced across the world). The fluidic teleportation canalso provide virtual replicated or multiple fluidic coupling, e.g. asingle device can be virtually fluidically connected to many“downstream” devices. Fluidic teleportation can also provide fortemporally distributed virtual fluidic coupling (e.g. the time lapsebetween the detection of the composition of a fluid exiting a firstdevice and the synthesis of a fluid with the same composition fordelivery to a second device (or many second devices) that is notfluidically coupled to the first device, can be very short, or verylong). More details, and exemplary implementations, of fluidteleportation are described below.

As noted above, fluidic teleportation can enable creation of a fluidicinterconnection between at feast two locations within a fluidic ormicrofluidic network without modifying the existing topology orarrangement a the network (e.g., without physically bridging orconnecting said locations by physical means). For example, the fluidicinterconnection can be established without connecting physical tubing orcreating a physical interconnection between the locations that allowdirect fluid flow. The interconnection can be established between atleast two discrete, separate, or disconnected fluidic networks (e.g.,chambers or components). The interconnection can be established betweenat least two sequential locations positioned opposite to the directionof a unidirectional fluid flow (e.g., the interconnection of a firstlocation to a second location that is upstream of the first location).The interconnection can be established between at least two locationsseparated by a flow restrictor, one-way valve, a filter, a componentthat selectively disallows the physical passage of chemical constituentsof the fluid, or combinations thereof.

A simple implementation of a fluidic teleportation system 600 isillustrated schematically in FIG. 9 . As shown, the fluidicteleportation system 600 includes a first fluidic synthesizer 610A, afirst microphysiological device 630A, a first multi-analyte fluidictransducer/biosensor 640A, and first sensor transmitter 650A. integratedinto a first device. As shown, the fluidic teleportation system 600further includes a second fluidic synthesizer 610B, a secondmicrophysiological device 630B, a second multi-analyte fluidictransducer/biosensor 640B, and a second sensor transmitter 650Bintegrated into a second device. As shown in FIG. 9 , the outflow of thefirst microphysiological device 630A can be coupled to the inlet of thesecond microphysiological device 630B without a direct fluidicconnection, by using a fluidic-to-digital signal transduction, a digitalsignal transmission, and then a digital-to-fluidic transduction, thuscreating virtual fluidic connection. As shown in FIG. 9 , multi-analytefluidic transducers/biosensors 640A located downstream of themicrophysiological devices 630A can transduce the fluidic composition ofthe effluent of microphysilogical device 630A into a digital signal thatcan be wirelessly transmitted by sensor transmitter 650A to the secondfluidic synthesizer 610B located upstream of the coupled secondmicrophysiological device 630B for reconstitution and subsequentperfusion.

Another implementation of a fluidic teleportation system 700 isillustrated schematically FIG. 10 . In the implementation, multiplemicrophysiological devices 730 are dynamically coupled by connectionsbetween the devices' respective multi-analyte transducers/biosensors 740and the sensor transmitters 750. Each of the microphysiological devices730 is fed via a fluid synthesizer 710. Since the system can beconnected through a fluidic-to-digital and subsequent digital-to-fluidictransduction, the devices can be arbitrarily distant from each other aslong as digital communication is possible.

FIG. 11A to are schematic illustrations of different topologies offluidic teleportation systems, according to various embodiments. Asshown in FIG. 11A, movement of a signal can be from a first device to asecond device. More specifically, the controller of Device 1 can controlthe fluidic synthesizer, the microphysiological device, and/or thebiosensor and data from Device 1 can be transmitted from the sensor datatransmitter of Device 1 to the controller of Device 2. Based on the datatransmitted from to the controller of Device 2 from the sensor datatransmitter of Device 1, the controller of Device 2 can modify itscontrol of the fluidic synthesizer, the microphysiological device,and/or the biosensor of Device 2. As shown in FIG. 11B, devices can bearranged in a serial virtual connection. In other words, data fromDevice 1 can be applied to the controller of Device 2 to control theoperation of Device 2 and data from Device 2 can be applied to thecontroller of Device 3 to control the operation of Device 3. As shown inFIG. 11C data from a first device can be applied in controlling theoperation of multiple additional devices in a multiplexed 1:2 (or 1:N)virtual connection. As shown in FIG. 11C, data front Device 1 is used incontrolling the operation of Device 2 and Device 3. As shown in FIG.11D, devices can be arranged in a feedback loop. In other words, outputfront a downstream device (e.g., Device 2) can be communicated to aninput of an upstream device (e.g., Device 1). In some embodiments, anyof the aforementioned topologies can be combined to create any desiredteleportation device. An example of a complex arrangement ofteleportation devices is shown in FIG. 12 .

The dynamic coupling between multiple microphysiological devices can beused, for example, for the assembly of a human-body-on-a-chip modelhaving a multitude of microphysiological devices that each recapitulatethe physiology or function of a particular human tissue, organ, orsystem, each connected to one or more other devices by digital fluidteleportation. This is illustrated schematically in an exampleembodiment in FIG. 12 . As shown in FIG. 12 , a system oforgan-on-a-chip microphysiological devices can include a gut device, aliver device, a kidney device, a bone device, a heart device, and a lungdevice. An oral drug can be administered (e.g. through a fluidicsynthesizer and fluid addressing system, and/or directly to the device,e.g. by a fluid handler, as described above, to the gut device and anaerosol drug can similarly be administered to the lung device. As shownin FIG. 12 , an oral drug can be simulated in an inlet to a gut device,and the gut device can communicate with a liver device, kidney device, abone device, and a heart device. Based on the data output from the gutdevice, the inputs to the liver device, the kidney device, the bonedevice, and the heart device can be controlled or manipulated. Based indata output from the liver device, the kidney device, the bone deviceand the heart device, the controller of the lung device can manipulatethe controller of the gut device. The operation and data derived formthe lung device can also be affected by the aerosol drug administered tothe lung device. In some embodiments, connections between the differentorgan devices can be virtual and/or physical. In some embodiment, thedifferent organ-on-a-chip devices can be contained on a singlemicrophysiological platform, or distributed across two or moremicrophysiological platforms. In some embodiments, a central controlsystem can control each of the organ devices. In some embodiments, thecentral control system can be integrated into a single apparatus thatincludes each of the organ devices, on a single microphysiologicalplatform or multiple microphysiological platforms. In some embodiments,one or more of the organ platforms can be implemented on removabledisks.

In some embodiments, digital fluid teleportation implementation (e.g.,from one organ platform to another organ platform) can include measuringthe concentration or presence of at least one target analyte in a firstfluid at a first location and synthesizing a second fluidic solutionusing a fluidic synthesizer. The second fluidic solution can include atleast one target analyte at the measured concentration or at aconcentration derived from mathematical adjustment or transformation ofthe measured concentration. The second fluid can be an identical, apartial, or a derivative mixture of the first fluid. In certainembodiments, the method can further include flowing the second fluidicsolution from the fluidic synthesizer to a second location withouttransporting the first fluidic solution from the first location to thesecond location through a continuous fluid flow. For example, a whole orpartial fluid composition characterized by at least one sensor at afirst location can be “digitally teleported” to a second location bywholly or partially reconstituting the first fluid at the secondlocation. These methods can be performed without requiring physicallyconnecting the first fluid at the first location to the second location.In certain embodiments, this method of digital fluid teleportation canallow an identical, partial, or derivative mixture of the first fluid atthe first location to be copied (by “digital fluid teleportation”)across any distance to one or a plurality of additional locations, solong as a means of data communication or data transport exists betweenthose at least two locations.

The method can include creating a first fluid mixture using a firstfluidic synthesizer, incubating a first microphysiological device withthe first fluid mixture to generate a second fluid mixture, incubatingthe first fluidic solution with a first microphysiological device at thefirst location to generate the second fluidic solution, measuring aconcentration of one or a plurality of target analytes in the secondfluid mixture using chemical sensors, and synthesizing a third fluidmixture using the first fluidic synthesizer to reconstituting anidentical, partial, or derivative mixture of the second fluid mixture.In non-limiting embodiments, the method can further include incubatingwith the third fluid mixture with the first microphysiological device,the second microphysiological device, at least two additionalmicrophysiological devices, or combinations thereof to generate a fourthfluidic solution. In non-limiting embodiments, the method can furtherinclude measuring a concentration of the at least one target analyte inthe fourth fluidic solution for subsequent perfusion.

In certain embodiments, the method for digital fluid teleportation canfurther include transmitting a measured concentration to at least oneexternal receiver. The external receiver can include at least one of acomputer, an electronic controller, an electronic control system, anetwork address, a network monitor, the first fluidic synthesizer, oneor a plurality of fluidic synthesizers, and a control server. Innon-limiting embodiments, the method for digital fluid teleportation canfurther include incubating a second tissue with the third fluidicmixture to generate a fourth fluidic mixture. In some embodiments, themethod can further include measuring a concentration of the targetanalyte in the fourth mixture for subsequent perfusion. In certainembodiments, digital fluid teleportation can enable the fluidicinterconnection of at least two microphysiological devices withoutrequiring the preemptive placement of said devices in a physicallyconnected manner or along a sequential fluid pathway. In certainembodiments, interconnections created by digital fluid teleportation canbe dynamically or discretely enabled, disabled, rerouted, or acombination thereof, continuously or at one or a plurality of intervals.

In certain embodiments, the disclosed subject matter provides methodsfor digital fluid teleportation between at least two organ-on-a-chipdevices. In certain embodiments, a fluidic chemical sensor or biosensorcan perform a real-time acquisition and readout of the composition of afirst fluid or fluid mixture at its location at a first organ-on-a-chipdevice. This readout can be transmitted digitally to be dynamicallyresynthesized and outflowed from a second location by a fluidicsynthesizer. The outflowing resynthesized fluid can be directed to thesecond organ-on-a-chip device including a fluid addressing system. Incertain embodiments, an identical, a partial, or a derivativeconstituent mixture of the first fluid can be delivered to the secondorgan-on-a-chip without the requirement that the first organ-on-a-chipbe connected to the second organ-on-a-chip. In such a manner by couplingone or more integrated fluidic biosensors with one or more fluidicsynthesizers capable of recreating the identical, partial, or derivativeconstituent mixture of the first sensor-characterized fluid mixture at asecond, different position on the chip, the first fluid can be “copied”from one position and “pasted” elseswhere on the same platform or ontoanother platform which possesses a fluidic synthesizer with theappropriate inlet compounds.

In certain embodiments, the sensor signal used for digital fluidteleportation can be digitally transmitted wirelessly, or through anytransmission network or protocol capable of transferring digital data.In certain embodiments, a digital signal can be transmitted back to afluidic synthesizer of the same organ-on-chin for modeling of feedbackloops.

In certain embodiments, digital fluid teleportation allows a fluidcomposition to be digitally transported in a manner, or to a location orlocations, that is incompatible with the limitations of physical fluidflow. For the purpose of illustration and not limitation, thetransportation can include transport across an air gap or other obstacleblocking fluid flow (e.g., a flow restriction or one-way valve),transport across a distance at a rate faster than can be achievedthrough certain physical fluid conduit (because a sensor signal can betransmitted at the speed of light), transport that is routed betweendestinations at a rate that exceeds the capacity of physical multi-wayvalving, or combinations thereof.

In certain embodiments, two physically discrete organ models can beconnected by the disclosed digital fluid teleportation method. As anon-limiting example, models of multi-organ systems can be formedbetween at least two microphysiological devices within themicrophysiological platforms that are integrated without physicalfluidic connections that link the microphysiological devices or organmodels directly. In certain embodiments, the disclosed system can beoperated without the unintended leakage of a fluidic biochemicalmicroenvironment between said microphysiological devices or fluidicdilution of constituents between the said organ models ormicrophysiological devices.

In certain embodiments, to connect a first microphysiological device toa second microphysiological device, the disclosed system can read thefluid composition with a biosensor for at least one fluidic analyte orconstituents at the outlet of the first microphysiological device,transmit the readouts to a fluidic synthesizer upstream of the secondmicrophysiological device to synthesize a second fluid mixture with anidentical, a partial, or a derivative constituent mixture of the firstfluid, and then flow this composition onwards to the secondmicrophysiological device. In non-limiting embodiments, a digital fluidteleportation method can be applied to transport one or a plurality offluids between any one or a combination of microphysiological devicesintegrated into the microphysiological platform shown in FIGS. 1 and 2 .The fluidic outflow of any microphysiological device can be sampled,reconstructed in the fluidic synthesizer, and addressed to any othermicrophysiological device by the fluid addressing system. This processcan be performed sequentially to link multiple microphysiologicaldevices. For example, a first microphysiological devices output can bereconstituted and be delivered to a second microphysiological device.The fluid outflow of the second microphysiological device can bereconstituted and delivered to a third microphysiological device, and soforth.

In certain embodiments, the connections established by digital fluidteleportation between two or more locations in the platform (e.g.,between microphysiological devices) can be newly established, modified,or closed in real-time, or in an otherwise dynamic manner. Unlike aninflexible system formed solely from physical fluid conduits, thedisclosed subject matter can allow coupling between microphysiologicalsystems after a stable state, a target state, or a desired operationalduration has been established. In non-embodiments, the coupling can bedisabled at predetermined time points. For example, the behavior of onemicrophysiological device can be evaluated in the temporary absence ofthe other. In certain embodiments, this dynamic establishment of fluidconnections by digital fluid teleportation can allow organ-on-a-chipmodels to be cultured in isolation for some target duration (e.g., oneday, or one week, or one month), after which they can be fluidicallycoupled to model organ-to-organ or tissue-to-tissue interactions. Incertain embodiments, this dynamic establishment of fluid connections bydigital fluid teleportation can allow two distinct microphysiologicaldevices to be cultured in isolation for some target duration (e.g., oneday, or one week, or one month). After the connection,microphysiological devices can be fluidically coupled to model dynamics(e.g., the exposure of a first biological entity to secretions orchemical compounds produced by a second biological entity, thebiological feedback phenomena produced from such a unidirectional orbidirectional coupling, or the biological reactions of a chain reactionbetween more than two biological entities).

In certain embodiments, the fluid characterized by one biosensor can befluidically resynthesized by digital fluid teleportation fortransmission multiple targets in order to produce dynamic fluidicduplications. For example, the fluid delivered to multiplemicrophysiological devices cultured as replicates (for purposesincluding, without limitation, statistical rigor or hypothesis testing)can be simultaneously coupled to the fluidic composition measured at theeffluent of a single driving microphysiological device. The effluent canbe characterized by a sensor placed at the outlet of said drivingmicrophysiological device. Then, upon teleportation to a fluidicsynthesizer, the fluidic synthesizer can reconstitute the mixture andflow an equal unit volume of the same mixture to each of the replicatemicrophysiological devices through a fluid addressing system. In certainembodiments, the replicates to which said fluid mixture is delivered canbe biologically identical. In certain embodiments, the replicates towhich said fluid mixture is delivered can be nominally biologicallyidentical (i.e., they can be cultured with the same initial conditionsfor purposes of being identical replicates), for predetermined purposes(e.g., classification or quantification of their resultant variability).In certain embodiments, the intentional variability can include a sourceof tissues cultured in different microphysiological devices, apathophysiological state of tissues cultured in said replicates, or theage of tissues cultured in said replicates. In certain embodiments, thistechnique can be used to analyze the different behaviors or responsesexhibited by healthy versus diseased organ models in response toexposure the same dynamic fluidic environment.

In certain embodiments, a fluid can be duplicated by digital fluidteleportation to two locations simultaneously, by the transmission ofthe sensor signal to two fluidic synthesizers. In certain embodiments, afluid can be duplicated by digital fluid teleportation to two locationssequentially by teleportation to one fluid synthesizer whose output isaddressed to a first location by a fluid addressing system. Then thefluid can be teleported to the same fluid synthesizer to address thesame fluid mixture as an output to a second location by the said fluidaddressing system. In certain embodiments, the desired fluid mixture canbe duplicated to three or more locations.

In certain embodiments, digital fluid teleportation can transportgreater or smaller volumes of a fluid composition to one or moresecondary locations relative to the volume of said fluid that is sampledby a plurality of sensors at a first location. For purposes ofillustration and not limitation, a sensor at a first location can samplea chamber containing 0.1 milliliters of fluid, and this measured fluidcomposition can be transported by digital fluid teleportation to asecond location at which 1.0 ml of an identical fluid composition can besynthesized by a fluidic synthesizer.

In certain embodiments, a sampled fluidic composition can be transportedby digital fluid teleportation not only spatially, but also throughtime, or temporally. In certain embodiments, temporal teleportation canbe achieved by placing a delay between one or more sensors at a firstlocation, and a fluidic synthesizer that recreates the mixture at asecond location. For purposes of illustration and not limitation, thiscan be utilized to mimic natural phenomena. For example, the naturalphenomena can include the transit time of a hormone or biological entitybetween tissues in the human body. When multiple microphysiologicaldevices are interconnected by digital fluid teleportation, thesequential transport of a metabolite or biological entity between aplurality of tissues or organs in a timed sequence can be performed.

In certain embodiments, the one or more sensors can perform a discreteor continuous recording of a fluidic composition at a first locationduring a first time period. The sensors can also digitally store thesensor data that composes the recording. The recording can be reloadedto reproduce the recorded composition through a fluidic synthesizer. Incertain embodiments, this method of loading a saved recording of a fluidcomposition can be used to recreate the fluid composition from saidrecording at a later date. In certain embodiments, this method of savingand then later loading a recording of a fluid composition can be used toarchive particular behaviors or dynamics that are observed in anexperiment in a microphysiological platform. The behaviors or dynamicscan be recreated at a later date on a different microphysiologicalsystem. In certain embodiments, the sensor readings from the duration ofan entire experiment can be saved, such that they can be accessed laterfor various purposes. The purpose can be a playback of said recording inwhole or in part through the real-time re-synthesis of said recordedfluid compositions at a fluidic synthesizer. In certain embodiments, thetechnique of saving sensor data that characterizes a discrete orcontinuous fluid composition can increase the experimentalreproducibility of multi-organ coupled systems for investigating rarephenomena.

FIG. 13 shows a flow chart of a nonlimiting example of a method 800 ofdigital fluid teleportation. The method 800 includes receiving dataindicative of a measured concentration of a target analyte in a firstfluidic solution at a first location as shown in 801. In someembodiments, the first location can be an organ-on-a-chip model. Themethod 800 further includes providing control instructions to a fluidicsynthesizer to cause the fluidic synthesizer to synthesize a secondfluidic solution that includes the target analyte at the measuredconcentration or at a concentration derived from mathematical adjustmentor transformation of the of the measured concentration and thatreconstitutes a full or partial composition of the first fluidicsolution as shown in 802. In some embodiments, the second location canbe on an organ-on-a-chip model. The method 800 further includes causingthe second fluidic solution to flow to a second location that isfluidically isolated from the first location, shown in 803.

In some embodiments, the method 800 can include causing the secondsolution to flow to an additional location (i.e., different from thesecond location), the additional location fluidically isolated from thefirst location. In some embodiments, the first location and the secondlocation can be on a single microphysiological platform. In someembodiments, the first location can be on a first microphysiologicalplatform and the second location can be on a second microphysiologicalplatform. In some embodiments, the first physiological platform and thesecond physiological platform can be engaged with a single controlsystem. In some embodiments, the control system can receive the data,provide the control instructions, and cause the second fluidic solutionto flow. In some embodiments, the first microphysiological platform canbe engaged with a first control system and the second microphysiologicalplatform can be engaged with a second control system. In someembodiments, the method can further include receiving second dataindicative of a measured concentration of the second target analyte in athird fluidic solution resulting from passage of the second fluidicsolution through the second microphysiological device, shown in 804. Insome embodiments, the method 800 can further include providing secondcontrol instructions to a second fluidic synthesizer to cause the secondfluidic synthesizer to synthesize a fourth fluidic solution thatincludes the second target analyte at the measured concentration of thesecond target analyte or at a concentration derived from mathematicaladjustment or transformation of the measured concentration of the secondtarget analyte and that reconstitutes a full or partial composition ofthe third fluidic solution, shown in 805. In some embodiments, themethod 800 can further include causing the fourth fluidic solution toflow to a third microphysiological device at a third locationfluidically isolated from the first location and the second location,shown in 806.

In some embodiments, a method of fluid teleportation can includemeasuring the constituent concentrations of dissolved chemical elementsand routes in a fluid by utilizing a sensor specific to a firstlocation. Optionally mathematical transformations can be applied to themeasured concentrations. The mathematical transformations can include,for purposes of illustration and not limitation, doubling or halving themeasured concentration of a solute during the production of a fluid at afluid synthesizer, adding together two or more signals, constituentconcentrations, or sets of constituents and their concentrations, addingone or more constant values to one or more constituent concentrations orsignals, or combinations thereof. The concentration can be transmuted toa control system or a remote system. A fluid mixture with equivalentchemical or solute concentrations can synthesized by mixing undilutedstock solutions together at a second location. In some embodiments, themixture of the stock solutions can include an identical, a partial, or aderivative constituent mixture of the first fluid's measuredconcentration. The fluid mixture can be delivered to a third location.In some embodiments, the third location can be discrete or physicallyseparated from the first location. In some embodiments, the thirdlocation can be discrete or physically separated from the secondlocation. In some embodiments, teleporting the solution contents fromthe second location to the third location can be without a continuousfluidic connection between the second location and the third location.

In non-limiting embodiments, the microphysiological platform can recorda rare stimulus in a microphysiological device through a plurality ofsensor modalities or targets, record said stimulus, and replicate inreal-time or at a later time on command. A plurality of such recordingscan be replicated through multiple fluidic synthesizers, which can belocated remotely from each other. In non-limiting embodiments, thesampled fluidic composition can be modified and teleported in real-timeor at a later time to a fluidic synthesizer. For example, theconcentrations of a plurality of constituent belonging to a measuredfluid composition that is being transported by digital fluidteleportation can be modified from the concentration of concentrationsmeasured in the fluidic recording. This technique can allow acharacterized outflow of a dynamic environment at a firstmicrophysiological device to be reconstituted at a secondmicrophysiological device without requiring the physical presence of thefirst microphysiological device in proximity and in fluid communicationwith the second microphysiological device. In certain embodiments, thefluidic environment generated by a first microphysiological device canbe resynthesized at a second microphysiological device at one of morelater time points without requiring the first microphysiological deviceto be physically recreate said measured environment local to the firstmicrophysiological device. Accordingly this technique can decrease cost,material requirements, and processing time.

In certain embodiments, the transduced digital signal of a chemical orbiosensor can be manipulated and adjusted before it is resynthesized asa part of digital fluid teleportation. In certain embodiments, a firstfluid composition can be resynthesized as a second fluid by it fluidicsynthesizer such that this second fluid is a partial or derivativemixture of the first fluid. In certain embodiments, the partial orderivative mixture of the first fluid can contain at least oneconstituent at concentrations measured in the first fluid. The partial.or derivative mixture of the first fluid can be resynthesized. in thesecond fluid at a different concentration froth the measuredconcentration. The different concentrations can result from manipulationof the measured concentration by processes including algorithmictransformation, mathematical transformation, or transformation by signalprocessing techniques. For example, the disclosed techniques can allowcharacterizing a subset constituents measured in the first fluid (e.g.,determined by the stock solutions available to the fluidic synthesizeror by the linear combinations of stock solutions available to thefluidic synthesizer), a subset of constituents measured in the firstfluid (e.g., determined by experimental requirements such as theincompatibility or toxicity of certain compounds at one location withthe biological entities in the culture at a second location), a supersetof constituents composed of a partial or whole mixture of theconstituents in the first fluid mixture, additional constituents notpresent or not measured in the first mixture, or combination thereof.

In certain embodiments, digital fluid platform of a partial orderivative subset of a first sampled fluid at a first location to createa second fluid at a second location can prevent the bulk transport ofcertain undesired dissolved compounds in the first fluid by bulk fluidflow to the second location. In certain embodiments, such digital fluidteleportation of a partial or derivative subset of a first sampled fluidcan prevent the bulk transport of certain undesired dissolved compoundswithout requiring a physical mechanism for selectively filtering saidundesired compounds from the bulk fluid mixture.

In certain embodiments, the microphysiological platform including aplurality of individual microphysiological devices to cultureorgan-on-a-chip models an utilize digital fluid teleportation capabilityto selectively transport only a partial or derivative subset of theconstituents in a first sampled fluid at a first organ-on-a-chip modelto a second location corresponding to a second organ-on-a-chip model. Incertain embodiments the capability can be utilized to prevent theunintended transport of dissolved compounds (e.g., growth factors,biological secretions, drugs, or chemical compounds) from a firstorgan-on-a-chip to a second organ-on-a-chip model. In some embodiments,this allows multiple organ-on-a-chip models to be combined into a “humanbody on a chip” configuration that features multiple tissue typescultured discrete organ-on-a-chip models (each housed in amicrophysiological device, within a microphysiological platform) thatare fluidically interconnected by digital fluid teleportation. In suchembodiments for multi-organ or “human body on a chip” configurations,the capacity to selectively transport only a partial or derivativesubset of the constituents in a sampled fluid can be utilized totransport only particular class chemical compounds (e.g., biologicallysecreted compounds, hormones, paracrine factors, endocrine factors,signaling molecules, or combinations thereof) without transportingundesired compounds (e.g., growth factors intended for the culture ofonly one tissue type metabolic waste products, undesired signalingmolecules, biological entities including viruses and bacteria,extracellular vesicles, or combinations thereof). In certainembodiments, the disclosed microphysiological platform can prevent thebulk leakage or transport of a biochemical microenvironmentcharacteristic from a first microphysiological device or organ-on-a-chipmodel to a second microphysiological device or organ-on-a-chip model.The disclosed microphysiological platform prevents the bulk leakage byfluidically synthesizing a mixture that includes the concentrations ofonly a targeted set of compounds in the fluidic effluent of a firstmicrophysiological device before introducing it to a secondmicrophysiological device or a sequence of microphysiological devices.Such selectivity over constituents transported in a fluid is notpossible in an alternative system that is configured to employ a directfluid connection in which whole fluid (or a volumetric subset of saidfluid) physically proceeds in bulk by pumping or directional flow in asequential manner between a plurality of biological cultures connectedin series.

In the devices, which possess direct fluid connections between two ormore sequential biological cultures, any dissolved constituents in thesequentially propagating fluid mixture including cannot be isolated orremoved without additional complex processes (e.g., chromatographicelution). Thus, the undesired constituents can flow sequentially throughall of the biological cultures. Such bulk, the non-selective flow offluid prevents the mimicry or reconstitution of physiological processes(e.g., the sequestration in the natural human body of certain localorgan-specific biochemical microenvironments, biochemical signalingpathways, growth factors, morphogens, paracrine signaling compounds, orother biochemical constituents to just one tissue or a spatial subset oftissues in parallel with large-scale or body-wide propagation ofcompounds such as endocrine or hormonal signaling mechanisms).

In certain embodiments, fluidic interconnects that are formed by digitalfluid teleportation between a plurality of microphysiological devicespermit the separation of short-range biological signaling ordevelopmental cues within one tissue from the biological signalsinherent to, secreted by, expected by, or required by other tissues,while still permitting selected signals to propagate by digital fluidteleportation, thereby overcoming limitations inherent to requiring onesingle “universal cell culture medium” that is compatible with allbiological tissues cultured in a microphysiological platform or system.

In certain embodiments, the microphysiological devices can constitutebiological entities. The microphysiological devices can includeorgan-on-a-chip models, a different growth medium used for each tissueor organ-on-a-chip model while superimposing the chemical signatures ofcompounds suspected of being involved in physiologically relevantorgan-to-organ signaling. For example or purposes of illustration andnot limitation, a microphysiological platform can use the fluidaddressing system to perfuse “pancreas-on-a-chip” firstmicrophysiological device with a first specific media formulation. Theplatform can sample the concentration of the insulin biologicallysecreted into the effluent of said media formulation by the firstpancreas-on-a-chip microphysiological device and reconstitute saidinsulin concentration into a second media formulation with a fluidicsynthesizer, and subsequently deliver this second media formulation to a“fat-on-a-chip” second microphysiological device. In certainembodiments, this process of selectively transporting a subset ofchemical constituents allows the effects of coupling specific biologicalcompounds to be studied without the rest of the fluidic environment alsoleaking between the organs. In certain embodiments, the concentration ofthe constituent or constituents in fluid transported by digital fluidteleportation can be derived from one or more of the following: aphysical measurement, an algorithmic manipulation of physicalmeasurement, a predicted value, or an arbitrary value.

In certain embodiments, the disclosed system can amplify the concentrateon of a plurality of constituents in a measured signal to overcomeproportionality limitations between accurate relative sizing of culturedbiological tissues. For example, when the physical volume or cellularquantity of a biological tissue cultured in a first microphysiologicaldevice is insufficient to recapitulate the physiological or desiredconcentration profiles of a specific chemical entity resulting frombiological secretion, biochemical modification absorption, metabolism,or other biological dynamic process affecting the concentration of thedissolved chemical compounds, the measured quantities of secreted orabsorbed product by the microphysiological device can be mathematicallytransformed of scaled up of down, linearly of nonlinearly of with alookup table, during the operation of the fluidic synthesizer to complywith or mimic a physiological range or the desired target range. Forexample, individual pancreatic beta cells can release the samequantities of insulin in a microphysiological device as they can in thehuman body. However, there can be a circumstance in which a smallerquantity of pancreatic islet beta cells to a pancreas-on-a-chipmicrophysiological device cannot secrete the same physiologicallyrelevant quantity of insulin in response to increase glucoseconcentrations in the media as the concentration produced in the humanpancreas. A fat-on-a-chip microphysiological device coupled directly orby digital fluid teleportation to the pancreas-on-a-chip device andexposed to said insulin cannot induce sufficient glucose uptake to causea significant change in the glucose content of the media. In thisexample, by amplifying both the detected concentration of secretedinsulin and the corresponding decrease in glucose in the media wheninstructing the fluidic synthesizer, the two organ-on-a-chipmicrophysiological devices can be coupled in a physiologically relevantmanner despite limitations to their size in certain embodiments of amicrophysiological device.

In certain embodiments, the disclosed system can recreate a specificcontinuous fluidic composition including a plurality of chemicalcompounds and their target concentrations as a function of time, at oneor more specified locations, and synthesize said fluidic compositionsimultaneously or sequentially to said locations. In certainembodiments, by leveraging digital fluid teleportation to pass fluidiccomposition data over communications networks including the internet,the disclosed system can facilitate the collaborative investigation ofmulti-tissue dynamics using resources and microphysiological (and themicrophysiological devices therein) that are housed in differentfacilities. In certain embodiments, multi-laboratory,multi-institutional, or even multi-national collaborative efforts can beachieved through the transduction and subsequent digital fluidteleportation of effluent composition from a microphysiological deviceinto a digital signal, the transmission of that signal to a plurality ofmicrophysiological platforms, and transduction of that signal to a fluidmixture using a fluidic synthesizer. Teams of experts in the distinctorgan or tissue systems can digitally interconnect their tissue modelstogether to form a human body model that spans the globe. For purposesof illustration and not limitation, a beating heart-on-a-chip in Francecan branch to perfuse a lung-on-a-chip model in Pennsylvania and akidney-on-a-chip at a laboratory in California, which can, in turn, becoupled to a liver model in Australia, and so forth.

As described above with references to FIGS. 2A and 2B, the disclosedsubject matter provides hardware-level components for operating andcontrolling the microphysiological platform and its components including(without limitation) microphysiological devices. These describedcomponents can be replaced, rearranged, modified, and reengineered toaccommodate any possible variations of the system.

FIG. 14 illustrates an exemplary system hardware architecture of anembodiment of a control system 970 operating with multiplemicrophysiological platforms 900 and various external devices andsystem. As shown, the control system 970 includes a main controller 972,web clients 973, a fluid synthesizer controller 974, an addressingsystem controller 976, biolines servers 977, a networked PC 979, a datareceiver 981, and an imager 982. As shown, the microphysiologicalplatforms 900 include fluidic synthesizers 910, fluid addressing systems920, microphysiological devices 930, and biosensors 940. Main controller972 can run fault-tolerant control system software that can, in certainembodiments, synchronize one or more of the following digital fluidteleportation, fluid coupling, data acquisition, fluidic control,pneumatic control, system monitoring, environmental monitoring,experimental feedback, user interaction, automatic responses to nominalor fault states, environmental climate control, data transmission, webserver operation, communication with additional microphysiologicalplatforms, and synchronization with additional microphysiologicalplatforms. In certain embodiments, the main controller 972 can serve aweb browser-based user interface to local users' computers, laptops,and/or portable devices. In certain embodiments, the user interface tothe microphysiological platform's control system can run in a webbrowser in order to be compatible with a variety of platforms, be theymobile or traditional desktops, as long as they have a web browser. Incertain embodiments, a web-based user interface can remove thebottleneck of requiring multiple users to share a single physicalcomputer (e.g., a master control system itself). In some embodiments,the core software can run on the main controller 972, and users can useor log in to lightweight user interfaces or user clients on their owndevices (e.g., via the web client 973). In certain embodiments andwithout limitation, users can utilize the user interface to do one ormore of the following: design new experiments, implement or sequence themethods required in new experiments, define microphysiological deviceexperiments, modify existing experiments, monitor acquired data, exportdata, export graphical elements or analyses including plots of data, andmanage digital fluid teleportation connections.

In some embodiments, fluidic synthesizer controllers 974 and/oraddressing system controllers 976 can include shift registers for portexpansion that can be used for the real-time control of the pneumaticvalves on the microphysiological devices 930. The fluidic synthesizercontrollers 974 and/or addressing system controllers 976 can includeindustrial microcontrollers that can drive the relays for valveactuation and motion control (e.g., the imager 982) in response tohigher-level control commands from the main controller 972. In someembodiments, the imager 982 can include an SPR imaging camera. In someembodiments, the imager 982 can include a laser scanner. For example,each microphysiological platform 900 can have 14 valves (each 4-port,2-position) for the fluid addressing systems 920 (7 on each side) and 48valves (each 3-port, 2-position) for the fluidic synthesizers 910 (1-24on each side). In non-limiting embodiments, each microphysiologicalplatform 900 can have 12 4-way/2-position valves plus 4 3-way/2-positionvalves; or 28 3-way/2-position valves. The latter configuration canallow the entire fluid addressing system 920 to be depressurized, whichcan allow all connected microphysiological devices 930 to be washedsimultaneously. To drive the fluidic synthesizer 910, themicrophysiological platform 900 can have as many 3-way/2-position valvesas there can be connected mixture compounds. The connected compounds canbe mixed at different rations, or mixed at the same ratio. In someembodiments, two inlet solutions to the fluidic synthesizer 910 that canbe mixed at the same ratio can be driven by one valve. In someembodiments, the fluidic synthesizers 910 can be expanded to accommodatemore than 25 distinct mixture inputs per side. Since the required numberof valves increases accordingly, latching shift registers can be chainedto create a parallel array control pins for the pneumatic valves.

In certain embodiments, the biosensors 940 at each organ-sensor unit canbe transduced to a digital signal by the imager 982. The imager 982 canbe connected to the networked PC 979, whose purpose is to transmit thistransduced data to the main controller 972. In some embodiments, thenetworked PC 979 can include a small computer. In some embodiments, thenetworked PC 979 can include a laptop. The main controller 972 can readthe output directly from the imager 982 when the additional real-timedata processing requirements do not cause any slowdown or interference.A connection from the local main controller 972 to software servers(e.g., biolines serve 977) connected to the internet, cloud, orsoftware-as-a-service (SaaS) system can be established for softwarelicense management, access to virtual tissue libraries, and real-timedigital interfacing with off-site organ devices. Multiplemicrophysiological platforms 900 can be operated locally by a singlemain controller 972. When interfacing with off-site microphysiologicalplatforms 900 and microphysiological devices 930 (platforms and devicesoperated by separate master control servers), data transmission can bemediated the internet or cloud software servers. These technologies canallow improved-quality and easy-to-establish connections that can bemaintained and monitored for events including, without limitation,connection dropouts or to predict bandwidth requirements, load balancingconsiderations, and data traffic routing optimization.

The disclosed subject matter provides software applications andtechnologies for operating the microphysiological platform 900. Thedisclosed biological virtualization technologies for statistical dataprocessing and modeling virtual tissues and virtual organs can beintegrated into a main controller 972 and be made configurable in a userclient. As a nonlimiting example, the disclosed system can have a maincontroller 972 which can provide a fault-tolerant software forsynchronizing and managing all operations of the microphysiologicalplatform 900 in real-time. The software of the main controller 972 candetermine pulse-width modulation (PWM) biases in fluidic synthesizers910 based on user settings and transduced signals from the biosensors940. As a nonlimiting example, the open-close cycling of valves for flowmetering in a manner controlled by pulse width modulation can allowproportional control over flow rates through microfluidic valves (e.g.,in the fluid addressing system 920 or the fluidic synthesizer 910). Incertain embodiments the PWM can be applied at a certain frequency topermit full opening and closing cycles in a valve. For example, where 0%pulse width can correspond to an always-closed valve and 100% pulsewidth can correspond to an always-open valve, a valve with a 60% pulsewidth can remain open for twice longer in each pulse cycle than a valvewith a pulse width. The rate of fluid flow can be controlled byadjusting the opening period. In certain embodiments, PWM can be appliedat a certain frequency to preclude the full actuation (closing oropening) of a valve between the signal-high and signal-low portions of apulse. For example, a sufficiently high PWM frequency can permit such anelastomeric valve to flutter or undulate closely about a steady-stateposition, whereby a valve with a 60% pulse width can maintain ssteady-state position that allows twice the flow of a valve with a 30%pulse width. In certain embodiments, the fluid-filled fluid controllines can be integrated into the valving the microphysiological platform900 and driven using syringe pumps or peristaltic pumps.

The software of the main controller 972 can control themicrophysiological platform 900 by transmitting commands tomicrocontrollers (e.g., the fluidic synthesizer controller 974 and/orthe addressing system controller 976) to activate specificmicrophysiological device 930 addresses in the fluid addressing system920 for monitoring flow rates, fluid levels, and pneumatic linepressures. In certain embodiments, the software of the main controller972 can control the microphysiological platform 900 to perform dataprocessing, host a local web server for web client logins, package datafor transmission to local web clients (e.g., a data receiver 981 a localdata repository, and local data servers), and maintain a connection toany one or more of data and control servers. In certain embodiments,National instruments LabView can be used for control softwareimplementation. In certain embodiments, other programming languages canbe used for control software implementation, including one or acombination of the following without limitation: C++, C#, Python, Java.

In certain embodiments, the control system 970 ran detect leaks,clogging, or a combination of leaks and clogging of themicrophysiological platform 900 by polling the data on an outlet flowmeter and alert users by email, text message, or other direct orindirect communications method. In certain embodiments, upon detectingleaks or clogging events, the immediate response of the control system970 can be configurable (e.g., suspend flow into the cloggedmicrophysiological device 930, suspend flow across the entire device930, pause the experiment, or continue as normal).

In certain embodiments, the control system 970 can provide user clientsoftware. For example, users of the control system 970 can design,monitor, and process experiments on a software client that can be servedby the main controller 972 or by an internet server and run in their webbrowser. In certain embodiments, the user client software can run as alocal application on a computer (e.g., the networked PC 979) or as adistributed application among a plurality of computers. In certainembodiments, the software can present aspects of the ongoing experiments(including, without limitation, portions of measured data) alongside aheads-up display of the current status of the fluid addressing system920 on the device, shown alongside the two columns of microphysiologicaldevices 930. In certain embodiments, users can click on individualmicrophysiological devices 930 to expand a settings box. In certainembodiments, each microphysiological device 930 can be customized (e.g.,named, given a distinctive icon, visual feature, visual characteristic,or a combination thereof). In certain embodiments, eachmicrophysiological device 930 can have all of its sensor 940 and flowproperties specified in the user interface. In certain embodiments,real-time data can be visualized in graphical form on a subpanel of theinterface allowing, an at-a-glance summary of a microphysiologicaldevice's status. In certain embodiments, said user interface can allowaccessory data to be imported and associated with a plurality of aspectsof existing data, including timestamps or sensor recordings.

In certain embodiments and for purposes of illustration and notlimitation, in response to seeing unexpected sensor readings, a user canimage a microphysiological device 930 in question, and then associatethat image as an accessory data file to the corresponding time points ofrecorded sensor or platform operation for later analysis. Innon-limiting embodiments, real-time data subpanel can be minimized. Incertain embodiments, digital fluid teleportation outline view canprovide a visualization of connections between microphysiologicaldevices 930 on a plurality of microphysiological platforms 900. Incertain embodiments, the design of the interface can improve thefunctionality and the clarity of the obtained data by providing a visualaesthetic in the form of bold colors. In certain embodiments the visualstyle of the user interface can complement the clarity of any presenteddata. Software produced by the disclosed subject matter can supportalternative-colors as well as improved-contrast schemes to permitaccessibility to colorblind or visually impaired users. In someembodiments, microphysiological devices 930 can be shown in a userinterface as movable blocks, and synthesized connectors can beestablished by dragging arid dropping visual links or ties between themovable blocks. In certain embodiments, experiments can be designed in auser interface using graphical elements including blocks, lines, orcurves. In certain embodiments, experiments can be designed byinteraction with the microphysiological platform and platform softwarethrough a command-line interface (CLI).

In certain embodiments, the disclosed subject matter provides theinternet-accessible software which can run on centralized softwareservers. In certain embodiments, the accessible internet software canmanage communications between multiple microphysiological platforminstallations, client licensing and accounts, and provides access to astandard library of templates, optimized protocols, and virtual tissuesas part of a subscription service. In certain embodiments, the disclosedsystem can operate across decentralized cloud platforms (e.g., Amazon'sAWS and/or Microsoft Azure).

Screen Forward Implementation

In some embodiments, the microphysiological platform and control systemarchitectures described above can function collectively as ahigh-throughput screening system. The screening system can define one ormore target conditions (e.g., chemical compounds, therapeutic drugstreatments and/or fluidic compositions) and identify responses ofcultured tissues in microphysiological devices (e.g., the time-dependentresponses and/or endpoint responses) which can result from the definedconditions. The screening process can be started from known conditionsor externally applied condition profiles (e.g., a drug concentrationprofile that mimics dosing with meals) to identify the endpoints thatresult from each such condition (e.g., a “screen-forward” approach). Anexample of such a screen forward approach is illustrated in the flowchart of FIGS. 15A and 15B, depicting method 1000. For example, as shownin FIGS. 15A and 15B, compounds, chemical concentration profiles,conditions, and/or physiological modifiers can be defined for biologicalscreening, shown in 1001. If an exemplary system includes asoftware-guided experiment design as determined in 1002, software-guidedstatistical analyses (e.g., power analysis, combinatorial or factorialanalysis, orthogonal experiment design. D-optimal experiment design,and/or experiment cost management) can be performed, shown in 1003.Certain experimental designs (e.g., quantity or placement of biologicalreplicates in microphysiological devices) can be determined without thesoftware-guided experiment design, shown in 1004. Then, based on theexperimental design, biological replicates (e.g., tissues, organ models,organ model systems, or a combination thereof) can be dispensed intomicrophysiological devices of the exemplary microfluidic system, shownin 1005. The biological replicates can be, immediately or after amaturation period, exposed to the pre-defined experimental conditions(e.g., certain concentrations of nutrients, drugs, bacteria, viruses,gene editing agents, or a combination thereof), shown in 1006.Integrated biosensors, if present and enabled, as determined in 1007 ofthe exemplary system can perform continuous or discrete sensing oftargets (e.g., DNA, RNA, proteins, growth factors, hormones), shown in1008. The detected results and can be stored and displayed on thegraphical user interface, shown in 1009. The biological replicates canbe cultured for the pre-determined period, shown in 1010. If integratedbiosensors are not determined to be present and enabled in 1007, 1008and 1009 are skipped and the method 100 proceeds to 1010. Upon reachingthe experiment endpoint period, endpoint measurements can be performedaccording to the experiment design, shown in 1011. For purposes ofillustration and not limitation, said endpoint measurements can includeone or a combination of the following: staining by immunohistochemistry(IHC), immunocytochemistry (ICC), tissue extraction, microscopy, RNA orDNA isolation, transcriptomics, proteomics and cell isolation. Themeasured endpoint data can be stored and reported to a user, shown in1012.

In non-limiting embodiments, the disclosed system can screen multipleconditions. Various dynamic and continuous conditions (e.g., a drugconcentration profile, and/or a pre-determined drug concentration basedon drug partition models) can be formulated in real-time by thedisclosed fluidic synthesizer. In certain embodiments, the disclosedsystem can test one or a combination of dynamic and continuousconditions for screening. As a nonlimiting example, the disclosed systemcan determine the effects of different concentrations of a vascularendothelial growth factor (VEGF) receptor inhibitor on tumorangiogenesis, the effects of dosing a VEGF inhibitor at differentfrequencies or concentrations on tumor angiogenesis, or the effects ofdosing said VEGF inhibitor at a dynamic concentration profile thatmimics metabolism and/or transport in a human body.

Screen Backward Implementation

In certain embodiments, the disclosed system can perform a screeningstudy or screening experiment by identifying known endpoints and findingcausative conditions that generate or produce said endpoints. Thedisclosed system can define a target endpoint state and search aplurality of initial conditions or condition profiles that can producethe target endpoint state through one or a succession of iterativerefinements (i.e., a “screen-backward” approach). For example, FIGS.16A-16D depict a method 1100, in which the target biological endpointtarget state, profile, performance, and/or morphology can be defined,shown in 1101. If an exemplary system can include a software-guidedexperiment design as determined in 1102, software-guided statisticalanalyses (e.g., power analysis, combinatorial analysis, orthogonalexperiment design, D-optimal experiment design, experiment costmanagement, experimental modifiers, drugs, therapeutic compounds,chemical profiles, and tissue compositions to define dimensions of thetest space) can be performed, shown in 1103. Certain experimentaldesigns (e.g., experimental modifiers, drugs, therapeutic compounds,chemical profiles, and/or tissue compositions) can be determined withoutthe software-guided experiment design, shown in 1104. Then, based on theexperimental design, biological replicates (e.g. tissues, organ models,organ model systems, or a combination thereof) can be dispensed intomicrophysiological devices of the exemplary microfluidic system, in1105. The dispensed biological replicates can be, immediately or after amaturation period, exposed to the pre-defined experimental conditions(e.g., nutrients, drugs, bacteria, viruses, and/or gene editing agentsat one or a range of concentrations), shown in 1106. If integratedbiosensors are present and enabled, as determined in 1107, the method1100 can include performing continuous or discrete sensing of targets(e.g., DNA, RNA, proteins, growth factors, hormones) at 1108. Thedetected results and data can be stored and displayed on the graphicaluser interface, shown in 1109. As shown in 1110, 1111, and 1112, themethod of a screen-backward mode includes determinations of whetherintegrated biosensors are present and enabled, whether the experimentalduration has been reached if defined), and if sensor-based feedback isenabled, respectively. If both sensor-based feedback and integratedbiosensors are enabled, and the experimental duration has not beenreached, the method 1100 includes utilizing, sensor measurements toperform feedback control of biological behavior by modifying cultureconditions, shown in 1113. This loop continues until the experimentalduration is reached. In non-limiting embodiments, the biosensors canprovide real-time feedback based on the detected data regardingexperiment duration and/or biological behavior. The defected data can beused to modify the culture conditions, shown in 1112. The replicates inmicrophysiological devices can be cultured for the pre-determinedperiod, shown in 1114. The method 1114 can occur once the experimentalduration has been reached or if integrated biosensors are not presentand enabled. Upon reaching the experiment endpoint period or phase,endpoint measurements can be performed according to the experimentdesign, shown 1115. The measured endpoint data can be stored andreported to a user, shown in 1116. If any replicates can attain targetendpoint state, shown in 1116, the test conditions required to attainthe target endpoint state can be presented to the user, shown in 1118.Otherwise, the disclosed system can repeat the screen-backward procedurein a sequential or iterative manner until the target endpoint state canbe attained via 1119, looping back to 1105. As a nonlimiting example,various test conditions can be iteratively defined for the next sequenceof experiments. The test conditions can be defined automatically orsemi-automatically with the software guidance, or by one or more manualinputs, requirements, and/or constraints.

In non-limiting embodiments, the disclosed system can define a statewith limited tumor angiogenesis and limited cell death from systemictoxicity, and then screen for the tine-dependent dosing condition a VEGEinhibitor that results in this intended endpoint. In non-limitingembodiments, the disclosed system can be used for biological screeningprocedures. For example, the system can be employed on tissues culturedfrom a reporter cell line of human pancreatic beta cells that expressesfluorescent protein during insulin secretion. The target endpoint statecan be defined as a specific time-dependent fluorescent intensityprofile (which can correlate to a time-dependent insulin releaseprofile) in response to the exposure to increased glucose concentrationin the fluid mixture. The disclosed system can interatively screen forthe combination and time-dependent profile of culture conditions (e.g.,from a set of growth factors, glucose, and drug compound precursorsolutions) that can produce this target endpoint response.

In certain embodiments, the disclosed screen-forward and/orscreen-backward modes can be selected for the multi-device ormulti-tissue models defined in microphysiological devices on onemicrophysiological platform or a plurality of connectedmicrophysiological platforms. In the screen-forward or screen-backwardmodes, the target states or target conditions can be defined formultiple systems. For example, the screen-forward or screen-backwardconditions or constraints can be applied for one tissue or for multipletissues. The conditions or constraints can be defined as an ideal targetrange for some biological behaviors or biochemical reactions. Innon-limiting embodiments, the conditions or constraints can be optimizedas a cost function relating to the health of one tissue in terms of theother as a ranking of priority. For example, a target end-state that canprevent the behavior in one tissue while minimizing effects of drugtoxicity in another tissue can be defined for the screen-forward and/orscreen-backward modes.

In certain embodiments, a user interface can be designed as shown forpurposes of illustration and not limitation in FIG. 17A and in FIG. 17B.In certain embodiments, the operational mode of the microphysiologicalplatform can influence the elements, layout, and functionality of theuser interface.

The disclosed subject matter provides systems and methods for developingand integrating “virtual tissues” in certain embodiments. When theresponses of a microphysiological device (e.g., microphysiologicaldevice 130, as described above with reference to FIG. 1 , a particularbiological entity or biological tissue) to a given input space ofpossible combinations of stimuli can be characterized, a physicalembodiment of that particular microphysiological device can be replacedwith a virtualized, simulated representation based upon predicted orpre-established behaviors or responses. For example, any of the devicesdescribed above with reference to FIGS. 11A to 11D and FIG. 12 caninclude virtual or simulated devices (e.g., virtual tissuerepresentations). In certain embodiments, this virtual device (e.g.,virtual tissue) is simulated computationally by making predictions orstatistical models for expected behavior based on data collected fromobservations of physical embodiments of the device being simulated(within microphysiological devices, in animals, in humans, as gatheredfrom the scientific literature, or a combination thereof). In certainembodiments, this virtual device is simulated computationally based onheuristics, rules, or datasets. In certain embodiments, this virtualdevice is simulated based on speculative predictions, the virtualdevice's digital outputs can be similar or identical to the outputsmeasured in sensors from the corresponding physical version for whichthe virtual device was substituted. Accordingly, such a virtual devicecan be coupled to one or a plurality of additional physicalmicrophysiological devices or additional virtual devices, and theresponses, absorptions/secretions, and behaviors of the first virtualdevice can be modeled computationally.

In certain embodiments, biological behavior of a target tissue can bepredicted by using a virtual tissue including by creating a statisticalpredictive model of a target tissue based on physical observations ofthe target tissue and applying the statistical predictive model topredict a modification of a first fluidic solution. The modification cancomprise a chemical or a biological change that is induced when thefirst fluidic solution is incubated with the target tissue to producethe second fluidic solution. In non-limiting embodiments, the firstfluidic solution can be an input of the virtual tissue, and the secondfluidic solution can be an output of the virtual tissue. In someembodiments, a first continuous mathematical function or continuousmeasurement can be used as the input to the virtual tissue and a secondcontinuous mathematical function can be generated as a correspondingoutput. For example, a continuous mathematical function can be acontinuous relationship with time. For purposes of illustration and notlimitation, a continuous input function to a virtual tissue for acompound “A” whose concentration increases linearly as a function oftime can be written as A(t)=A₀ t, where “A₀” is a constant defining therate of accumulation; if said virtual tissue secretes a compound “B”such that compound “B” is always maintained at twice the concentrationof “A,'” then the virtual tissue's continuous output function over forcompound “B” would be B(t)=2 A(t), which can also be written as B(t)=2A₀ t. In certain embodiments, symbolically linking the input and outputsof the virtual tissues in this manner allows these equations to becalculated rapidly though numeric or analytical solvers, to produce asimulated outcome. In certain embodiments, the output data can begenerated by utilizing an optional mathematical transformation numericalvalues produced by the virtual tissue output.

In certain embodiments, predicting a biological behavior of a targettissue using a virtual tissue can further include coupling at least twovirtual tissues by connecting the output of a first virtual tissue tothe input of a subsequent virtual tissue. In non-limiting embodiments, anetwork of at least three virtual tissues can be established. Eachvirtual tissue can receive at least one input and generate at least oneoutput. Each of the at least one output can be connected to zero or atleast one virtual tissue inputs. In non-limiting embodiments, each ofthe at least one input and output can be mathematically transformedbefore or after connecting to a virtual tissue. The at least one inputand output data for each virtual tissue can be recorded, eitherdiscretely or continuously over some duration of time within the scopeof the experiment.

In certain embodiments, predicting a biological behavior of a targettissue using a virtual tissue can further include identifyingconstituents of the second fluidic solution calculated by the virtualtissue output and synthesizing a fluidic solution based on theidentified constituents using fluidic synthesizer.

In certain embodiments, the responses, absorption/secretions, andbehaviors of multiple virtual tissues can also be modeledcomputationally in said fashion. In certain embodiments, theobservations can be derived from existing data in the scientific ormedical literature. In certain embodiments, the observations can bederived from measurements taken in a microphysiological platform. Theobservations can be used to predict the behavior of a biological entity(that, in certain embodiments, can be cultured in a microphysiologicaldevice), and allow for the biological entity to be simulated as avirtual tissue rather than physically deployed within microphysiologicaldevice. These models can range from being computationally simple (e.g.,a virtual tissue can be embodied by a lookup table that maps theconcentration of a single input factor or chemical concentration to acorresponding quantity of a second compound to secrete) to beingcomputationally complex models based on statistical algorithms used inmachine learning (see FIG. 18 ). In certain embodiments, for purposes ofillustration and not limitation, the virtual tissue can be simulated bya neural network, by a decision tree, by an ensemble method of aplurality of machine learning methods, or by statistical inference. Thetransduced digital signal from a biosensor associated with a realbiological entity being cultured in a microphysiological device can bethe virtual tissue's input or a subset of the virtual tissue's inputs,and the virtual tissue's output can, in turn, be used as the digitalinput to the fluidic synthesizer that delivers the media mixture(containing the secretions of the virtual tissue) to the tissue beingphysically cultured. In certain embodiments, a plurality of virtualtissues can be coupled or linked to a plurality of virtual tissues.

For example, as shown in FIG. 18 , an organ-on-a-chip model cultured ina microphysiological device can be characterized by obtainingsubstantial amounts of data through the utilization of amicrophysiological platform under a wide variety of culture conditions.Subsequently, based upon the collected data that maps input conditionsto phenotype, behavior, and secreted factors and compounds, the dynamicresponses of the cultured tissue in a given condition can becomputationally predicted. In certain embodiments, the prediction methodcan use the interpolation of existing data to generate a prediction. Incertain embodiments, machine learning or statistical learning techniquescan be employed to generate a prediction. By replacing real tissue witha virtual tissue and digitally interfacing it to the fluidic inputs ofreal tissues' fluidic synthesizers or other virtual tissues' digitalinputs, tissue virtualization can be realized. Accordingly, thedisclosed virtual tissue techniques can allow the expansion oraugmentation of microphysiological devices with additional virtualtissues in order to permit a full body-on-a-chip analysis for every realtissue in culture (e.g., by simulating all other tissues or organs in afull-body model).

In certain embodiments, the disclosed virtual tissue system canstatistically model real microphysiological devices based on measuredsensor or imaging data in real-time as they are running. In certainembodiments, the virtual tissue system can use said measured sensor orimaging data to display predictive extrapolations of their behavior tousers. In certain embodiments, when local microphysiological devices areengaged in digital fluid teleportation connections to off-sitemicrophysiological devices in multi-facility collaborations, continuousstatistical modeling, and machine learning algorithms can be used toprovide transient virtual tissues as stand-ins during temporary dropoutsof a data connection.

In certain embodiments, the method for predicting a biological behaviorof a target tissue or a target multi-tissue interaction using a virtualtissue can further comprise coupling the virtual tissue to amicrophysiological device by delivering the composition of the virtualtissue's output solution as a synthesized fluidic solution to themicrophysiological device. In certain embodiments, a measured partial orfull composition of fluid outflowing from a microphysiological devicecan be used as the data input to a virtual tissue. In non-limitingembodiments, at least one virtual tissue can be coupled with at leastone microphysiological device by delivering the synthesized fluidicsolution to the at least one microphysiological device through a fluidaddressing system. In some embodiments, the method can further includecreating a mixed network of microphysiological devices and virtualtissues by the coupling at least one virtual tissue with at least onemicrophysiological device. For example, multiple virtual tissues and/ormultiple real tissues in microphysiological devices within amicrophysiological system can be coupled by digital fluid teleportation,where “fluid” teleported to or from the virtual tissue is processedsolely as a digital signal (as a result of there being no physicaltissue at which a fluid analog can be delivered for interaction). Whenmultiple varieties of tissue cultures in a microphysiological platformcan be characterized to the same extent, multiple virtual tissues andfor multiple real tissue units can be coupled in the same mannerpreviously described for systems comprising solely physical (“real”)microphysiological devices cultures.

FIG. 19A illustrates an exemplary coupling of physicalmicrophysiological devices via digital transmission. FIG. 19B depicts anexemplary coupling a single physical microphysiological device to adynamic system of multiple virtual tissues. The virtual tissue canreceive the digitally transduced contents of fluid effluent from thereal microphysiological device, and process the data according to thecharacterized behavior of the biological entity that it is virtualizing.Then, the virtual tissue can transmit the corresponding effluent'sdigital-fluidic output to the fluidic synthesizer of the real tissue,where it can be reconstituted into a physical mixture. In non-limitingembodiments, as shown in FIG. 19C, a fully virtual system can beestablished by interfacing two or more virtual tissues together. Incertain embodiments, the screen-forward and screen-backward operationalmodes can be used for virtual tissue models to substitute certainphysical models in the disclosed system. In certain embodiments, ascreen-forward or screen-backward approach consisting exclusively ofvirtual tissues can be used to identify the most promising conditions orpredicted outcomes of a set of experiments. In certain embodiments, saididentified predictions can be used to select physical experiments to beconducted on real microphysiological devices.

In certain embodiments, the method for predicting a biological behaviorof a target tissue using a virtual tissue can further include generatingbehavioral data or observational data from the at least onemicrophysiological device. The behavioral or observational data can beused for improving the statistical predictive modeling. In non-limitingembodiments, the target tissue can be incubated with the at least onemicrophysiological device in a predetermined condition to provide thebehavioral or observational data for improving the statisticalpredictive model. In some embodiments, at least one component ofbehavioral or observational data can be provided to train thestatistical predictive model. In certain embodiments, at least onevirtual tissue can be used for predicting the biological behavior of thetarget tissue.

In certain embodiments, the feedback data can be obtained using thedisclosed biosensors (e.g., the biosensors 140, as described above withreference to FIG. 1 ). In some embodiments, the biosensors can beintegrated on-chip (i.e. on a substrate, such as substrate 101, asdescribed above with reference to FIG. 1 ). In some embodiments, thebiosensors can be detachable. For example, the disclosed system canfully function without biosensors. An exemplary system that does notpossess any biosensors can still utilize the fluidic synthesizer toformulate, in real-time, a fluid or a fluid mixture from one or morefluid reagents connected to the system. For example, there can bemultiple precursor fluid inlet connections from which a compositesolution can be formulated. The quantity of the connections can beexpanded based on necessity. An exemplary system without biosensors candeliver this fluid formulation to a target culture chamber by utilizingthe fluid addressing system to select the target chamber as the intendeddestination for the fluid. The fluidic synthesizer can modify thecomposition of this fluid in real-time to create a dynamic fluidformulation. In some embodiments, output data from the devices can bedelivered to a sensor external to the substrate (e.g., an “off-chip”sensor). In some embodiments, the output data can be sent to a vialeffluent line and/or a fluid handling system to sample fluid downstreamof the devices and deliver the fluid to the biosensor.

In certain embodiments, an exemplary system without integratedbiosensors can generate microliter quantities of a specified fluidicmixture to perform conditional or combinatorial screening on selectedchambers. Instead of using an integrated biosensor, an effluent fluidcan be sampled from the system's outlet ports, and the endpoints used toquantify the results of the screened conditions can be measured from thesampled fluid. For example, the endpoints measurements can includemeasurements of biochemical secretion, metabolism, biochemicalprocesses, brightfield imaging of the contents of the culture chambers,fluorescent imaging of the contents of the culture chambers,electrophysiological measurements (e.g., transepithelial electricalresistance (TEER) measurements), genomic/transcriptomic/proteomicquantification of the cells or subsets of cells in the culture chambers,retrieval of the cells for subsequent processing (e.g., paraffinsectioning or processing for electron microscopy), or a combinationthereof.

In certain embodiments, exemplary system without biosensors can record afluidic composition on an external biosensor platform and digitallyteleport the recorded fluidic composition to a fluidic synthesizer. Forpurposes of illustration and not limitation, the secretions of tissuecan be sampled into multiple discrete collections over the course of atime series, and the concentrations of the secretions can be measuredusing conventional assay techniques (e.g., by ELISA). Following themeasurement of these samples, the resultant concentration profile can beused as a fluidic composition, and thus such a recording can bereplicated through at least one fluidic synthesizer, which can belocated remotely from each other. In non-limiting embodiments, thesampled fluidic composition can be modified and teleported to a fluidicsynthesizer. As a nonlimiting example, the concentration of theteleported fluidic composition can be modified from the concentrationmeasured in the fluidic recording.

In certain embodiments, an exemplary system without biosensors canperform the screen-forward and/or screen-backward screenings. Theinitial conditions (whether fixed in screen-forward orheuristically/iteratively generated in screen-backward approaches) canbe formulated by the fluidic synthesizer and do not require sensorfeedback. Similarly, the endpoints for either mode can be chosen suchthat quantification can be produced off-chip. In non-limitingembodiments, the biosensors can be integrated into the disclosed systemfor collecting more data related to biological characterization and/orstrengthening of associated virtual tissue models.

In certain embodiments, the virtual tissue can reduce the physicalcomplexity of the microphysiological platform (relative to the use of anequivalent real/physical model) in exchange for increased computationalcomplexity. By incorporating virtual tissues as substitutes formicrophysiological devices, a scientific experiment or investigationconducted with the subject matter can be less expensive andtime-consuming. For example, the virtual tissues can be free frombiological contamination risks, and the virtual tissues do not requirebiological supplementation with culture media or growth factors.Furthermore, statistical models of virtual tissues can bedeterministically repeatable. In some embodiments, the virtual tissuecan operate in a training mode. In such a training mode, themicrophysiological platform can utilize physical microphysiologicaldevices to challenge one or more tissues in real-time as dictated by thestatistical uncertainty of the virtual tissue, with the goal being toimprove the predictive efficacy of the virtual tissue in regimes orcircumstances where its existing predictive capacity is weakest or mostprone to error. In certain embodiments, these regimes or circumstancescan include without limitation a set of multiplepreviously-unencountered fluid compositions in order to measure theresponse of the real tissues and thereby populate any lacking areas ofits statistical observation space. The disclosed training mode cansubject real tissues to edge cases and contrive useful circumstances inorder to strengthen the virtual model through machine learningapproaches. The disclosed training mode can ensure that the virtualtissue does not interpolate too far from physically derived data duringreal experiments, so as to maximize its predictive power and minimizeerror.

In certain embodiments, an experiment conducted by the disclosed subjectmatter can consist entirely of virtual tissues for predictive modeling.As a nonlimiting example, the application of the disclosed biologicalvirtualization can be used for both screen-forward and screen-backwardcomputational solvers. In certain embodiments, the disclosed virtualtissue system can answer exemplary predictive questions includingwithout limitation: “given these initial conditions, what tissuedynamics can be predicted to be observed over four weeks?” or “giventhese desired dynamical properties of a tissue, what formulation ofinitial conditions can be predicted to produce said properties?” Incertain embodiments, the disclosed computational technology can beutilized in pharmaceutical development for both drug candidate screening(forward-computation of drug effects, as a nonlimiting example, “howwill the influence of this drug change tissue behavior?”) well as drugcandidate identification (back-computation of desired drug targets as anonlimiting example “given that a progression to a healthy state overfour weeks is dependent on these initial computed changes in cellbehavior, what type of drug would cause these changes?”).

In certain embodiments, the disclosed subject matter provides abrowsable virtual tissue library which can be accessible from the usersoftware client during configuration. The virtual tissue library canprovide access to various virtual tissue modules. Furthermore, thevirtual tissue library can offer an interface to real tissues that canbe cultured in microphysiological devices, winch can be seeded andcultured at a central location and interfaced with a client's localtissue culture by off-site fluidic teleportation withclient-configurable seeding dates. Accordingly, the virtual library canbe a cheaper alternative to a user than continuously incubating theirown large library of cells or tissues in tissue cultures and managingthe cost of flasks, growth media, cell-line orders, and so forth.

In certain embodiments, the disclosed subject matter provides methodsfor connecting a microphysiological device to a virtual tissue. Anexample method can include characterizing a fluidic solution bymeasuring a concentration of at least one target analyte with a sensorat a microphysiological device, generating an input to the virtualtissue based on the measurement, and providing the input to the virtualtissue.

Tissue Culturing Use Cases

In certain embodiments, the disclosed subject matter can provide asystem for creating large quantities of functional tissues.Microphysiological platforms (e.g., microphysiological platform 100, asdescribed above with reference to FIG. 1 ) described herein can be usedfor therapeutic tissue engineering by creating targeted quantities oftissue for transportation. Microphysiological platforms can create thetissues at a scale that produces sufficient tissue mass for culturingfunctional tissues. For example, a fluidic synthesizer (e.g., thefluidic synthesizer 110, as described above with reference to FIG. 1 )can control the biochemical environment of multiple devices (e.g.,tissue culture chambers) producing tissues with low morphologicalvariability. Fluid addressing systems (e.g., fluid addressing systems120, as described above with reference to FIG. 1 ) can utilize fluidinputs to support a large quantity of microphysiological devices (e.g.,hundreds, thousands, or more) at consistent now rates, for consistentflow durations, and with consistent fluid composition.Microphysiological platforms described herein can allow functionaltissues to be cultured at an improved scale relative to traditionaltissue culture methods involving manual pipetting or deployment oftissue culture in the limited footprints of well plate systems.

In certain embodiments, the disclosed subject matter can provide asystem for differentiating and maturing tissues. For example, amicrophysiological platform can be formulated such that the entiredifferentiation process of organoid tissues can be automated by thefluidic synthesizer and fluid addressing system. The fluidic synthesizerand fluid addressing system can produce and deliver the requiredsolutions for differentiation to the target devices. The requiredsolutions can be a biological growth medium that can include variouscompounds for tissue differentiation (e.g., growth factors,transcription inhibitors, and/or nutrients). In non-limitingembodiments, the disclosed screen-forward and screen-backwardoperational modes can be used to optimize organoid growth,differentiation, and biological function. In some embodiments, theintegrated biosensors (e.g., the biosensors 140, as described above withreference to FIG. 1 ) can provide continuous feedback regarding thestate of the organoid. The integrated biosensors can allow afeedback-based differentiation protocol that can dynamically modify theculturing conditions. In some embodiments, the disclosed system also canprovide a fixed condition for developing organoid tissues.

Other Use Cases

In certain embodiments, the disclosed system can be used to evaluate theeffects of genetic modifications on tissues. As a nonlimiting example,the fluidic synthesizer and the fluid addressing system can be used toproduce a culture medium with agents that can modify, introduce, orknock out certain genes of cells. The gene editing agents can includevarious components that can be used in various gene-editing techniques(e.g., CRISPR-Cas9, TALEN, Meganucleases, Zinc finger, and/or genetherapies). In non-limiting embodiments, the disclosed system can beused to assay the effects of genetic modifications at the tissue scale.In certain embodiments., the disclosed system can deliver agents thatcan perform genetic modification to a targeted subset or subpopulationof cells. The integrated biosensors can monitor the status of thetissues and provide feedback in real-time.

In certain embodiments, the disclosed system can be used to delivercells or bacteria to the tissues in culture chambers. The fluidicsynthesizer can add living entities to the fluid mixture, which can bedelivered by the fluid addressing system. The disclosed system withliving entities in the fluid mixture can be used to screen biologicaltissues. For example, the effect of bacterial infection on lung tissuescan be screened by delivering one or multiple types of bacteria to oneor more target tissues cultured within the device. In non-limitingembodiments, the disclosed system can be used to screen immunotherapies.As a nonlimiting example, certain modified human cells including T-cellsaugmented with a chimeric antigen receptor (i.e., CAR-T cells) can flowto one or more tissues in culture chambers on the device. Bindingefficacy of different CAR-T cells to certain tissues, cancers, cells,drugs, and/or living entities can be assessed. In some embodiments,certain cancer tissues can be cultured in the microphysiologicaldevices, and immunotherapies including CAR-T cell therapies can bescreened for efficacy in targeting the cancer tissues, using either thescreen-forward or screen-backward operational modes.

In certain embodiments, the disclosed system can deliver viruses to thetissues cultured therein in microphysiological devices. The fluidicsynthesizer can add certain viruses to the fluid mixture, which can bedelivered by the fluid addressing system. The disclosed system withcertain viruses can be used to assay the susceptibility of the culturedtissues to the viruses. In non-limiting embodiments, the system caninclude vaccines and be used to assay the efficacy of vaccines or otherpreventative agents against viral inflection.

In certain embodiments, the disclosed system can deliver certain gasesto cultured tissues in microphysiological devices through the fluidaddressing system. The disclosed system with gases delivered to thetissues can be used to model embolism and/or human airway tissues. In noembodiments, certain gases can be included in a fluid mixture and bedelivered to one or more microphysiological devices. In someembodiments, gas compositions can be designed to mimic hypoxic ofhyperoxic conditions. In certain embodiments, a lung model can besubjected to hypoxic or hyperoxic conditions, and its resultant behaviorcan be examined by the disclosed system. In non-limiting embodiments,certain toxic gases can be delivered to tissues cultured in the culturechambers for assessing their effects on, or damage to, the tissue.

The disclosed subject matter provides systems and methods for physicallymodeling, computationally modeling, of a combination thereof to form amicrophysiological platform. The multiple microphysiological devices canbe interconnected. In certain embodiments, the biological tissues,organisms, or systems being interconnected can span numerous scales ofbiological and population-level entities. For example, the multipleinterconnected biological tissues or systems can model the interactionof different cell types, different biological organs, differentbiological organisms (e.g., to model disease transmission between twohumans), different species (e.g., to model infection of human tissue bya culture of bacteria); different biological scales (e.g., to modeleffects of metabolite secretion from a culture of one cell type oncultures of tissues from a population of human donors) or combinationsthereof.

In certain embodiments, the disclosed microphysiological devices can bespatially patterned to connect to at least one microfluidic channel offluidic connection to form a microphysiological platform or a subset ofa microphysiological platform. In certain embodiments, the at least onemicrofluidic channels can be used to deliver fluids to at least onemicrophysiological device spatially patterned within said microfluidicchannel or microfluidic channel network. For example, the fluids caninclude cell culture media, nutrients, glucose, amino acids, biologicaltherapeutics, drugs, drug products, toxins, gases, dissolved solids,dissolved chemical compounds, aerosolized compounds, biologically actingchemical fixatives, or combinations thereof. In non-limitingembodiments, the fluids can be delivered in a plurality of discreteperiods. In certain embodiments, the fluids in a predetermined range ofdoses can be delivered doses to the microphysiological device. Incertain alternative embodiments, the fluids can be deliveredcontinuously to the microphysiological device.

In certain embodiments, chemical compositions of a fluid delivered tothe microphysiological platform can require a modification. Themodification can be made based on biological experimentation (e.g., tomaintain homeostasis, or in response to sensor readings). In certainembodiments, the modification to the fluid's chemical composition can bedynamic and continuous. In non-limiting embodiments, the modificationcan be discrete or can be performed stepwise.

In certain embodiments, the disclosed fluidic synthesizer can beincluded in a microphysiological platform to synthesize fluid of adesired chemical composition. The desired chemical composition can besynthesized by mixing or combining a predetermined concentration ofdiscrete ingredients, partitioned fluids, inlet solutions, reagents, ora combination thereof corresponding to the desired composition. Innon-limiting embodiments, the synthesized fluid mixture can be deliveredto a plurality of microphysiological devices/platforms using a deliverymechanism (e.g., microfluidic channel or a fluid conduit).

The disclosed subject matter provides systems and methods for producinga “body-on-a-chip” system or a system composed of multipleinterconnected “organ-on-chip” models using a plurality ofmicrophysiological devices that are not in physical fluid communicationor are not physically fluidically accessible. In certain embodiments,digital fluid teleportation can fluidically interconnectmicrophysiological devices or microphysiological platforms betweendifferent laboratories, different institutions, or scientists withdifferent experimental capabilities.

It will be understood that the foregoing is only illustrative of theprinciples of the present disclosure, and that various modifications canbe made by those skilled in the art without departing from the scope andspirit of the present disclosure.

1-53. (canceled)
 54. A method for predicting a biological behavior of atarget tissue, comprising: applying one or more first inputs to astatistical predictive model to predict the biological behavior of thetarget tissue, wherein the statistical predictive model is created basedon one or more second inputs provided to a cultured tissue within atleast one microphysiological device and observational data of thecultured tissue acquired by one or more sensors, and wherein thecultured tissue is the same tissue type as the target tissue.
 55. Themethod of claim 54, wherein the observational data include one or moreof phenotypic data, morphological data, metabolic data, genotypic data,and proteomic data.
 56. The method of claim 54, wherein theobservational data include a composition of a fluid output from thecultured tissue and a biological composition of the cultured tissueafter exposure to a fluid input.
 57. The method of claim 54, wherein thecultured tissue is incubated within the at least one microphysiologicaldevice in a predetermined condition based on the one or more secondinputs.
 58. The method of claim 54, wherein the one or more secondinputs and the observational data are provided to train the virtualtissue model.
 59. The method of claim 54, wherein the observational datais used to assess biological efficacy or safety of at least one compoundor condition on humans or animals.
 60. The method of claim 54, whereinthe observational data is used for drug development or screening. 61.The method of claim 54, wherein the statistical predictive modelcomprises a virtual tissue model.
 62. The method of claim 54, whereinthe statistical predictive model comprises one or more of a neuralnetwork, a decision tree, and statistical inference.
 63. The method ofclaim 54, wherein applying the one or more first inputs to thestatistical predictive model to predict the biological behavior of thetarget tissue further comprises applying a mathematical transformationto numerical values produced by the statistical predictive model. 64.The method of claim 54, wherein the one or more sensors comprise atleast one of an imager and a biosensor.
 65. The method of claim 64,wherein the biosensor comprises an analyte sensor.
 66. The method ofclaim 64, wherein the imager is configured to collect image informationfrom the at least one microphysiological device.
 67. The method of claim64, wherein the imager is configured to perform at least one of brightfield imaging, optical microscopy, fluorescence microscopy, magneticresonance imaging, and computed tomography (CT) scanning.
 68. The methodof claim 64, wherein the biosensor comprises a chemical reaction orbioreaction recognition element configured to interact with the targetanalyte to produce a measurement of quantity or concentration of thetarget analyte, a recording system configured to record the measurementand an activity of the microphysiological device.
 69. The method ofclaim 54, wherein the target tissue includes one or more of a lungtissue, a bone marrow tissue, a bone tissue, a pancreatic tissue, anendocrine islet tissue, a liver tissue, a kidney tissue, a placentatissue, an eye tissue, an intestinal tissue, a bladder tissue, a braintissue, a mouth tissue, a tongue tissue, a tooth tissue, a nose tissue,a thymus tissue, a lymph node tissue, a lymphatic system tissue, athroat tissue, a specific human tissue, a specific human tissueundergoing a specific routine behavior, a lung tissue that is cyclicallybreathing, a specific human tissue undergoing an atypical condition, alung tissue undergoing an asthma attack, a specific human tissueundergoing a specific interaction with an outside agent, a lung tissuebeing infected with bacteria, a lung tissue exposed to environmentalfactors, a lung tissue exposed to pollution, a lung tissue exposed tocorrosive gas, a specific human tissue undergoing a specific interactionwith an outside agent that is intended for use as a therapeutic, aspecific human tissue undergoing a specific interaction with a drug, aspecific human tissue undergoing a specific interaction with abiological antibody, a specific human tissue undergoing a specificinteraction with a cellular therapy, or a lung tissue undergoing anasthma attack while being monitored for its interaction with abronchodilator as therapy for asthma.
 70. The method of claim 54,wherein the one or more second inputs comprise one or more of acomposition of a fluid input to the cultured tissue, a biologicalcomposition of the cultured tissue before exposure to the fluid input,and predetermined incubation conditions.
 71. The method of claim 70,wherein the composition of the fluid input to the cultured tissuecomprises one or more of a predetermined culture medium, a chemicalstimulus, and a biological stimulus.
 72. The method of claim 70, whereinthe predetermined incubation conditions comprise one or more oftemperature, a duration of incubation, and a gas composition.
 73. Themethod of claim 70, wherein the one or more second inputs are defined bya continuous function.
 74. The method of claim 54, further comprising:coupling the statistical predictive model to the cultured tissue withinthe at least one microphysiological device; applying one or more thirdinputs to the statistical predictive model to predict the biologicalbehavior of the cultured tissue; and delivering, based on the predictedbiological behavior of the cultured tissue, a synthesized fluidicsolution as the fluid input to the cultured tissue within the at leastone microphysiological device, wherein the one or more third inputs arebased on the cultured tissue within the at least one microphysiologicaldevice.
 75. The method of claim 74, further comprising: delivering thesynthesized fluidic solution to the cultured tissue within the at leastone microphysiological device through a fluid addressing system.
 76. Themethod of claim 54, wherein the biological behavior of the target tissuecomprises phenotypic behavior, genotypic behavior, and proteomicbehavior.
 77. The method of claim 54, wherein the statistical predictivemodel of the target tissue is based on off-chip observational dataincluding one or more of measurements of biochemical secretion,metabolism, biochemical processes, electrophysiological measurements,and genomic, transcriptomic, and/or proteomic quantification of cells ora subset of cells in the cultured tissue.
 78. A system, comprising: acontrol system comprising a controller configured to: apply one or morefirst inputs to a statistical predictive model to predict the biologicalbehavior of the target tissue, wherein the statistical predictive modelis created based on one or more second inputs provided to a culturedtissue within at least one microphysiological device and observationaldata of the cultured tissue acquired by one or more sensors, and whereinthe cultured tissue is the same tissue type as the target tissue. 79.The system of claim 78, wherein the at least one microphysiologicaldevice comprises materials configured for optical imaging.
 80. Thesystem of claim 78, wherein the cultured tissue within the at least onemicrophysiological device is an organ-on-a-chip model.
 81. A method forpredicting a biological behavior of a target tissue using a statisticalpredictive model, comprising: acquiring the statistical predictive modelfrom a browsable statistical predictive model library; and applying oneor more first inputs to the statistical predictive model to predict thebiological behavior of the target tissue, wherein the statisticalpredictive model is based on one or more second inputs and observationaldata acquired by one or more sensors of a cultured tissue within atleast one microphysiological device, and wherein the cultured tissue isthe same tissue type as the target tissue.
 82. A method of creating astatistical predictive model, comprising: providing one or more firstinputs to a cultured tissue within at least one microphysiologicaldevice; acquiring observational data from the cultured tissue; andcreating the statistical predictive model by correlating the firstinputs and the observational data.